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• An important transcription elongation facilitator is FACT (facilitates chromatin transcription) – 2 subunits: • Spt16 binds to H2A-H2B dimers acid-rich C-terminus is essential for these nucleosome remodeling activities • SSRP1 binds to H3-H4 tetramers – Facilitates transcription through nucleosome by promoting loss of at least one H2A-H2B dimer
• Acts as histone chaperone promoting re-addition of H2A-H2B dimer to nucleosome that has lost dimer
FACT (facilitates chromatin transcription)
MCM Mini Chromosome Maintenance complex, or Minichromosome Maintenance protein complex or mini-chromosome maintenance (MCM) 2-7 helicase complex has a role in both the initiation and the elongation phases of eukaryotic DNA replication, specifically the formation and elongation of the replication fork. MCM is a component of the pre-replication complex, which is a component of the licensing factor. MCM consists of six related polypeptides that form a ring structure.
Replication fork: When replicating, the original DNA splits in two, forming two "prongs" which resemble a fork,
background
CPT: Camptothecin
DNA topoisomerase I inhibitor
Result
genome-wide short hairpin RNA (shRNA) screen
A Genetic Screen for Proteins that Confer Resistance to Replisome
Stress
Candidate Gene Validation
MMS22L Is Required for Resistance to a Vari-ety of DNADamaging Agents
MMC: mitomycin C
MMS22L Interacts with NFKBIL2, FACT, and MCMs
Comparative Proteomic Analysis Software Suite (CompPASS) which employs an unbiased methodology for the identification of high-confidence candidate interacting proteins (HCIPs) from parallel non-reciprocal proteomic data and facilitates functional dissection of interaction networks.
MMS22L Interacts with NFKBIL2, FACT, and MCMs
NFKBIL2 Is Required for Resistance to DNADamaging Agents
MMS22L and NFKBIL2 Promote Ho-mologous
Recombination
MMS22L and NFKBIL2 Promote Ho-mologous
Recombination
MMS22L and NFKBIL2 Suppress Spon-taneous
DDR Activation
MMS22L Depletion Delays Recoveryfrom CPT-Induced Damage
Increased 53BP1 and H2AX Foci upon Lossof MMS22L or NFKBIL2
Increased 53BP1 and H2AX Foci upon Lossof MMS22L or NFKBIL2
Increased 53BP1 and H2AX Foci upon Lossof MMS22L or NFKBIL2
CTIP-Dependent Phosphorylation of RPA2 and RPA2
Foci Formation Accompany MMS22L-NFKBIL2Depletion
replication protein A (RPA)
CTIP-Dependent Phosphorylation of RPA2 and RPA2
Foci Formation Accompany MMS22L-NFKBIL2Depletion
CTIP-Dependent Phosphorylation of RPA2 and RPA2
Foci Formation Accompany MMS22L-NFKBIL2Depletion