Ccdc124 A novel protein involved in cytokinesis
Uygar H Tazebay
Gebze Technical University
Department of Molecular Biology and Genetics
Gebze Turkey
International Conference on Science and Technology YTSAM October 2016
Cytokinetic Abscission
Midbody An organelle to be exploredhellip
The position of midbody in dividing cells
Blockage of abscission in response to Cep55 knock-down
Ccdc124 is encoded by a conserved eukaryotic gene
CLUSTAL W (182) multiple sequence alignment
human -MPKKFQGENTKSAAARARRAEAKA---AADAKKQKELEDAYWKDDDKHVMRKEQRKEEK 56
zebrafish -MPKKFQGENSKSATARARKAEAKA---VADARKQKELEDALWEDNDKRVVKKEQRKDDK 56
Celegans -MPKKFASENPKVTAARDRKATAKK---DEADKKAKATEDAKWVDNDKLNNRKMQRKEDD 56
Anidulans MGGKKGGGENSKKAAGNARKAEAAANKKAIEDQKRAAEEDKQWAKGAK----SSSKKEEA 56
human EKRRLDQLERKKETQRLLEEEDSKLKGGKAPRVATSSKVTRAQIEDTLRRDHQL-REAPD 115
zebrafish ERKRLEALERKRENQRLLEEEDSKIKGKQTK--EGPSKVTRAQIEETLQSKQNV-KEIK- 112
Celegans EKKREEALRRKEENRKLAEEEMSSLGNKKPAG-AATQKVTRAHIHIRKEDEERINRELEE 115
Anidulans EAKKAEAARKKAERDALLAAEEASQPSKPKNNKSAAKKNAPSRGTLNLD-------QLDD 109
human TAEKAKSHLEVP---LEENVNRRVLEEGSVEARTIEDAIAVLSVAEEA-ADRHPERRMRA 171
zebrafish --EKEKSHLDVP---LEENVNRIVPEEGTVEARTIEDAIAVLSTKED--LDRHPERRMKA 165
Celegans KRKQEAQKIEVAGDLLVENLNKLEVEEG--EARNVDDALKVLGEEKALDDDKHPEKRMRA 173
Anidulans APSSRASALNAS------------------ GIDNALDALSLTSKDTSK-VDRHPERRYKA 150
human AFTAFEEAQLPRLKQENPNMRLSQLKQLLKKEWLRSPDNPMNQRAVPFNAPK-------- 223
zebrafish AYTAFEEANMPRVKMENPNMRLSQLKQQLKKEWTKSPENPLNQRAASYNTK--------- 216
Celegans AYLAFEEARLPELKLNHPTFRLSQLKQILKKEWQKSPENPLNARLLALNS---------- 223
Anidulans AYAAFEARRLPEIEAENPGLRRQQRIELVKKEFDKSPENPFNQVHVAFDASKEEIAAVRE 210
human ------------
zebrafish ------------
Celegans ------------
Anidulans AERKKTEARLTR 222
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Cytokinetic Abscission
Midbody An organelle to be exploredhellip
The position of midbody in dividing cells
Blockage of abscission in response to Cep55 knock-down
Ccdc124 is encoded by a conserved eukaryotic gene
CLUSTAL W (182) multiple sequence alignment
human -MPKKFQGENTKSAAARARRAEAKA---AADAKKQKELEDAYWKDDDKHVMRKEQRKEEK 56
zebrafish -MPKKFQGENSKSATARARKAEAKA---VADARKQKELEDALWEDNDKRVVKKEQRKDDK 56
Celegans -MPKKFASENPKVTAARDRKATAKK---DEADKKAKATEDAKWVDNDKLNNRKMQRKEDD 56
Anidulans MGGKKGGGENSKKAAGNARKAEAAANKKAIEDQKRAAEEDKQWAKGAK----SSSKKEEA 56
human EKRRLDQLERKKETQRLLEEEDSKLKGGKAPRVATSSKVTRAQIEDTLRRDHQL-REAPD 115
zebrafish ERKRLEALERKRENQRLLEEEDSKIKGKQTK--EGPSKVTRAQIEETLQSKQNV-KEIK- 112
Celegans EKKREEALRRKEENRKLAEEEMSSLGNKKPAG-AATQKVTRAHIHIRKEDEERINRELEE 115
Anidulans EAKKAEAARKKAERDALLAAEEASQPSKPKNNKSAAKKNAPSRGTLNLD-------QLDD 109
human TAEKAKSHLEVP---LEENVNRRVLEEGSVEARTIEDAIAVLSVAEEA-ADRHPERRMRA 171
zebrafish --EKEKSHLDVP---LEENVNRIVPEEGTVEARTIEDAIAVLSTKED--LDRHPERRMKA 165
Celegans KRKQEAQKIEVAGDLLVENLNKLEVEEG--EARNVDDALKVLGEEKALDDDKHPEKRMRA 173
Anidulans APSSRASALNAS------------------ GIDNALDALSLTSKDTSK-VDRHPERRYKA 150
human AFTAFEEAQLPRLKQENPNMRLSQLKQLLKKEWLRSPDNPMNQRAVPFNAPK-------- 223
zebrafish AYTAFEEANMPRVKMENPNMRLSQLKQQLKKEWTKSPENPLNQRAASYNTK--------- 216
Celegans AYLAFEEARLPELKLNHPTFRLSQLKQILKKEWQKSPENPLNARLLALNS---------- 223
Anidulans AYAAFEARRLPEIEAENPGLRRQQRIELVKKEFDKSPENPFNQVHVAFDASKEEIAAVRE 210
human ------------
zebrafish ------------
Celegans ------------
Anidulans AERKKTEARLTR 222
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Midbody An organelle to be exploredhellip
The position of midbody in dividing cells
Blockage of abscission in response to Cep55 knock-down
Ccdc124 is encoded by a conserved eukaryotic gene
CLUSTAL W (182) multiple sequence alignment
human -MPKKFQGENTKSAAARARRAEAKA---AADAKKQKELEDAYWKDDDKHVMRKEQRKEEK 56
zebrafish -MPKKFQGENSKSATARARKAEAKA---VADARKQKELEDALWEDNDKRVVKKEQRKDDK 56
Celegans -MPKKFASENPKVTAARDRKATAKK---DEADKKAKATEDAKWVDNDKLNNRKMQRKEDD 56
Anidulans MGGKKGGGENSKKAAGNARKAEAAANKKAIEDQKRAAEEDKQWAKGAK----SSSKKEEA 56
human EKRRLDQLERKKETQRLLEEEDSKLKGGKAPRVATSSKVTRAQIEDTLRRDHQL-REAPD 115
zebrafish ERKRLEALERKRENQRLLEEEDSKIKGKQTK--EGPSKVTRAQIEETLQSKQNV-KEIK- 112
Celegans EKKREEALRRKEENRKLAEEEMSSLGNKKPAG-AATQKVTRAHIHIRKEDEERINRELEE 115
Anidulans EAKKAEAARKKAERDALLAAEEASQPSKPKNNKSAAKKNAPSRGTLNLD-------QLDD 109
human TAEKAKSHLEVP---LEENVNRRVLEEGSVEARTIEDAIAVLSVAEEA-ADRHPERRMRA 171
zebrafish --EKEKSHLDVP---LEENVNRIVPEEGTVEARTIEDAIAVLSTKED--LDRHPERRMKA 165
Celegans KRKQEAQKIEVAGDLLVENLNKLEVEEG--EARNVDDALKVLGEEKALDDDKHPEKRMRA 173
Anidulans APSSRASALNAS------------------ GIDNALDALSLTSKDTSK-VDRHPERRYKA 150
human AFTAFEEAQLPRLKQENPNMRLSQLKQLLKKEWLRSPDNPMNQRAVPFNAPK-------- 223
zebrafish AYTAFEEANMPRVKMENPNMRLSQLKQQLKKEWTKSPENPLNQRAASYNTK--------- 216
Celegans AYLAFEEARLPELKLNHPTFRLSQLKQILKKEWQKSPENPLNARLLALNS---------- 223
Anidulans AYAAFEARRLPEIEAENPGLRRQQRIELVKKEFDKSPENPFNQVHVAFDASKEEIAAVRE 210
human ------------
zebrafish ------------
Celegans ------------
Anidulans AERKKTEARLTR 222
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
The position of midbody in dividing cells
Blockage of abscission in response to Cep55 knock-down
Ccdc124 is encoded by a conserved eukaryotic gene
CLUSTAL W (182) multiple sequence alignment
human -MPKKFQGENTKSAAARARRAEAKA---AADAKKQKELEDAYWKDDDKHVMRKEQRKEEK 56
zebrafish -MPKKFQGENSKSATARARKAEAKA---VADARKQKELEDALWEDNDKRVVKKEQRKDDK 56
Celegans -MPKKFASENPKVTAARDRKATAKK---DEADKKAKATEDAKWVDNDKLNNRKMQRKEDD 56
Anidulans MGGKKGGGENSKKAAGNARKAEAAANKKAIEDQKRAAEEDKQWAKGAK----SSSKKEEA 56
human EKRRLDQLERKKETQRLLEEEDSKLKGGKAPRVATSSKVTRAQIEDTLRRDHQL-REAPD 115
zebrafish ERKRLEALERKRENQRLLEEEDSKIKGKQTK--EGPSKVTRAQIEETLQSKQNV-KEIK- 112
Celegans EKKREEALRRKEENRKLAEEEMSSLGNKKPAG-AATQKVTRAHIHIRKEDEERINRELEE 115
Anidulans EAKKAEAARKKAERDALLAAEEASQPSKPKNNKSAAKKNAPSRGTLNLD-------QLDD 109
human TAEKAKSHLEVP---LEENVNRRVLEEGSVEARTIEDAIAVLSVAEEA-ADRHPERRMRA 171
zebrafish --EKEKSHLDVP---LEENVNRIVPEEGTVEARTIEDAIAVLSTKED--LDRHPERRMKA 165
Celegans KRKQEAQKIEVAGDLLVENLNKLEVEEG--EARNVDDALKVLGEEKALDDDKHPEKRMRA 173
Anidulans APSSRASALNAS------------------ GIDNALDALSLTSKDTSK-VDRHPERRYKA 150
human AFTAFEEAQLPRLKQENPNMRLSQLKQLLKKEWLRSPDNPMNQRAVPFNAPK-------- 223
zebrafish AYTAFEEANMPRVKMENPNMRLSQLKQQLKKEWTKSPENPLNQRAASYNTK--------- 216
Celegans AYLAFEEARLPELKLNHPTFRLSQLKQILKKEWQKSPENPLNARLLALNS---------- 223
Anidulans AYAAFEARRLPEIEAENPGLRRQQRIELVKKEFDKSPENPFNQVHVAFDASKEEIAAVRE 210
human ------------
zebrafish ------------
Celegans ------------
Anidulans AERKKTEARLTR 222
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Blockage of abscission in response to Cep55 knock-down
Ccdc124 is encoded by a conserved eukaryotic gene
CLUSTAL W (182) multiple sequence alignment
human -MPKKFQGENTKSAAARARRAEAKA---AADAKKQKELEDAYWKDDDKHVMRKEQRKEEK 56
zebrafish -MPKKFQGENSKSATARARKAEAKA---VADARKQKELEDALWEDNDKRVVKKEQRKDDK 56
Celegans -MPKKFASENPKVTAARDRKATAKK---DEADKKAKATEDAKWVDNDKLNNRKMQRKEDD 56
Anidulans MGGKKGGGENSKKAAGNARKAEAAANKKAIEDQKRAAEEDKQWAKGAK----SSSKKEEA 56
human EKRRLDQLERKKETQRLLEEEDSKLKGGKAPRVATSSKVTRAQIEDTLRRDHQL-REAPD 115
zebrafish ERKRLEALERKRENQRLLEEEDSKIKGKQTK--EGPSKVTRAQIEETLQSKQNV-KEIK- 112
Celegans EKKREEALRRKEENRKLAEEEMSSLGNKKPAG-AATQKVTRAHIHIRKEDEERINRELEE 115
Anidulans EAKKAEAARKKAERDALLAAEEASQPSKPKNNKSAAKKNAPSRGTLNLD-------QLDD 109
human TAEKAKSHLEVP---LEENVNRRVLEEGSVEARTIEDAIAVLSVAEEA-ADRHPERRMRA 171
zebrafish --EKEKSHLDVP---LEENVNRIVPEEGTVEARTIEDAIAVLSTKED--LDRHPERRMKA 165
Celegans KRKQEAQKIEVAGDLLVENLNKLEVEEG--EARNVDDALKVLGEEKALDDDKHPEKRMRA 173
Anidulans APSSRASALNAS------------------ GIDNALDALSLTSKDTSK-VDRHPERRYKA 150
human AFTAFEEAQLPRLKQENPNMRLSQLKQLLKKEWLRSPDNPMNQRAVPFNAPK-------- 223
zebrafish AYTAFEEANMPRVKMENPNMRLSQLKQQLKKEWTKSPENPLNQRAASYNTK--------- 216
Celegans AYLAFEEARLPELKLNHPTFRLSQLKQILKKEWQKSPENPLNARLLALNS---------- 223
Anidulans AYAAFEARRLPEIEAENPGLRRQQRIELVKKEFDKSPENPFNQVHVAFDASKEEIAAVRE 210
human ------------
zebrafish ------------
Celegans ------------
Anidulans AERKKTEARLTR 222
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 is encoded by a conserved eukaryotic gene
CLUSTAL W (182) multiple sequence alignment
human -MPKKFQGENTKSAAARARRAEAKA---AADAKKQKELEDAYWKDDDKHVMRKEQRKEEK 56
zebrafish -MPKKFQGENSKSATARARKAEAKA---VADARKQKELEDALWEDNDKRVVKKEQRKDDK 56
Celegans -MPKKFASENPKVTAARDRKATAKK---DEADKKAKATEDAKWVDNDKLNNRKMQRKEDD 56
Anidulans MGGKKGGGENSKKAAGNARKAEAAANKKAIEDQKRAAEEDKQWAKGAK----SSSKKEEA 56
human EKRRLDQLERKKETQRLLEEEDSKLKGGKAPRVATSSKVTRAQIEDTLRRDHQL-REAPD 115
zebrafish ERKRLEALERKRENQRLLEEEDSKIKGKQTK--EGPSKVTRAQIEETLQSKQNV-KEIK- 112
Celegans EKKREEALRRKEENRKLAEEEMSSLGNKKPAG-AATQKVTRAHIHIRKEDEERINRELEE 115
Anidulans EAKKAEAARKKAERDALLAAEEASQPSKPKNNKSAAKKNAPSRGTLNLD-------QLDD 109
human TAEKAKSHLEVP---LEENVNRRVLEEGSVEARTIEDAIAVLSVAEEA-ADRHPERRMRA 171
zebrafish --EKEKSHLDVP---LEENVNRIVPEEGTVEARTIEDAIAVLSTKED--LDRHPERRMKA 165
Celegans KRKQEAQKIEVAGDLLVENLNKLEVEEG--EARNVDDALKVLGEEKALDDDKHPEKRMRA 173
Anidulans APSSRASALNAS------------------ GIDNALDALSLTSKDTSK-VDRHPERRYKA 150
human AFTAFEEAQLPRLKQENPNMRLSQLKQLLKKEWLRSPDNPMNQRAVPFNAPK-------- 223
zebrafish AYTAFEEANMPRVKMENPNMRLSQLKQQLKKEWTKSPENPLNQRAASYNTK--------- 216
Celegans AYLAFEEARLPELKLNHPTFRLSQLKQILKKEWQKSPENPLNARLLALNS---------- 223
Anidulans AYAAFEARRLPEIEAENPGLRRQQRIELVKKEFDKSPENPFNQVHVAFDASKEEIAAVRE 210
human ------------
zebrafish ------------
Celegans ------------
Anidulans AERKKTEARLTR 222
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 has high identity and similarity rates among eukaryotes
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
582351222Aspergillus nidulans
650389238Oryza sativa
737438213Drosophila melanogester
726504223Caenorhabditis elegans
891700216Danio rerio
910728217Xenopus laevis
955888217Mus musculus
987955223Canis familiaris
996991223Pongo pygmaeus
100100223Homo sapiens
Similarity IdentityAmino acid
LengthAlignment
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
1 2
3
4 5
6 7 8 9 10 11
12 13 14
Characterization of Ccdc124
at the protein level
220
160
120
100
90
80
70
60
50
40
30
25 20
220
160
120
100
90
80
70
60
50
40
30
25 20
Mw kDa
Mw kDa
FLAG-Ccdc124 EV
Ccdc124-FLAG
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Prophase Metaphase Anaphase
Ccdc124
DNA
Merge
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 is a novel centrosome protein
Ccdc124 DAPI Aktin (Pha)
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Centrosome structure
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124
g-tubulin
Interphase Prophase Metaphase Anaphase
DNA
Merge
Cytokinesis
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 g-tubulin Merge + DNA
Subcellular localization of Ccdc124 during cytokinetic abscission
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 is a novel midbody protein
Ccdc124 DAPI Aktin (Pha)
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Cytokinetic abscission is blocked when Ccdc124 is knocked-down by RNAi
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Genomic copies of Ccdc124 mutated in HEK-293 by CRISPRCas9 technique
Ccdc124 loci on Chromosome 19
24 nucleotide (8 aa) genomic deletion
91 nucleotide genomic deletion (Frameshift mutation)
No protein
Mut 215 aa
WT 223 aa
CRISPRCas9 mutated copies
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Multi-nuclear cells in response to Ccdc124 mutations by CRISPRCas9
Ccdc124 g-tubulin DNA
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Enlarged aberrant Ccdc124 +- cell phenotypes
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Time-lapse imaging of aberrant cell formation
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Collective cell migration in development and epithelial mesenchymal transition
Modeling and analysis of collective cell
migration in an in vivo three-dimensional
environment
Caia et al 2015 PNAS USA 113(5)E2134-
2141
Epithelialndashmesenchymal transition (EMT) is essential during embryo development and found in common pathologies such as organ fibrosis and in the initiation of metastasis for cancer progression
In vivo collective cell migration requires an
LPAR2-dependent increase in tissue fluidity
Kuriyama et al 2014 Journal of Cell Biology 206 (1)113-121
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
New (fair) city border proposed by Uygar
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Acknowledgements
Hani Alotaibi (IBG-DEU) Pelin Telkoparan (YIU) Elif Yaman (Leica)
Merve Tuzlakoğlu Oumlztuumlrk (GTUuml) Oumlmer Guumllluumlluuml (GTUuml) Sinem Guumll (GTUuml) Oumlzge Arslan Eltan (GTUuml)
Batu Erman (SU) Fred Wittinghofer (Dortmund) Johannes L Bos (Utrecht) Lars Meijer (Utrecht)
Turkish Scientific and Technological Research Council (TUumlBİTAK) 114-Z-349
Turkish Ministry of Development 2014-KB-101977
Feyzi Akkaya Scientific Research Funds (FABED)
EMBO Short Term Fellowships Program
European Science Foundation (ESF) ndash Funct Genomics
People Agencies and Foundations
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
bullCellular polarization
bullCell-cell adhesion
bullCell-ECM attachment
bullCellular migration
Rap2 is involved in
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Interaction Partners of Ccdc124
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Interaction Partners of Ccdc124
bull Fusion of biotin ligase to Ccdc124
anti-Ccdc124
b-Actin
35
55
70
35
55
Wb anti-Myc Antibody
myc-BirA myc-BirA-Ccdc124
Bio- Bio+
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Anti-MYC Anti-g-TUBULIN MERGED + DNA
Myc
-Bir
A-C
cdc1
24
bull Monitoring cellular localization of Myc-BirA-Ccdc124
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
LC-MSMS analysis of Myc-BirA-Ccdc124 biotinylated cellular factors
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Interpretation of initial MS data
~2000 proteins (total number of proteins)
330 protein (unique to Ccdc124-Bioid after NT and EV-Mock corrected)
212 strong candidates after assessing uniquetotal peptide ratios
(ge 020)
58 priority interactors of Ccdc124 after comparing previous MS data
Validations by co-IP experiments
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
RNA processing Cell division and cytoskeleton component
Others
Viral life cycle primary cilium assembly regulation of gene expression protein complex subunit organization etc
Candidate interaction partners of Ccdc124
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 interacts with components of RNA maturation and decay
STAU2-
Flag +
pcDNA
31
STAU2-
Flag +
pMEX-
HA
STAU2-
Flag +
CCDC12
4-HA
Untrans
fected
CCDC12
4-HA +
pcDNA
31
CCDC12
4-HA +
3X Flag
CMV10
STAU1-
Flag +
pcDNA
31
STAU1-
Flag +
pMEX-
HA
STAU1-
Flag +
CCDC12
4-HA
MR
250
130
100
70
55
35
25
STAU2-
Flag +
pcDNA
31
(SUP)
STAU2-
Flag +
pMEX-
HA
(SUP)
STAU2-
Flag +
CCDC12
4-HA
(SUP)
Untrans
fected
(SUP)
CCDC12
4-HA +
pcDNA
31
(SUP)
CCDC12
4-HA +
3X Flag
CMV10
(SUP)
STAU1-
Flag +
pcDNA
31
(SUP)
STAU1-
Flag +
pMEX-
HA
(SUP)
STAU1-
Flag +
CCDC12
4-HA
(SUP)
MR
Supernatant
250
130
100
70
55
35
25
IP anti-HA WB anti-Flag
Staufen2 functions in Staufen1-mediated mRNA decay by binding to itself and its paralog and promoting UPF1 helicase
but not ATPase activity Park E Gleghorn ML and Maquat LE Proc Natl Acad Sci U S A 2013 Jan 8110(2)405-12 doi
101073pnas1213508110 Epub 2012 Dec 20
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 a new RNA metabolism protein
(2014)
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
Ccdc124 is ubiquitously expressed in human tissues
CCDC124
b-actin
GAPDH
[kb]
10
![Natural Water Chemistry - unizg.hr6].pdf · KISELE KIŠE: 22 22 3 s aq aq 3 aq aq 2 g l CaCO 2H Ca H CO Ca CO H O Utjecaj na ljudske tvorevine: → otapa vapnenac → uzrokuje koroziju](https://static.documents.pub/doc/80x56/5e46c115a8e98a0d2a58b630/natural-water-chemistry-unizghr-6pdf-kisele-kie-22-22-3-s-aq-aq-3-aq-aq.jpg)
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