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www.reactome.orgMurcia - 3 February 20111
A Database of human biological pathways
Bijay Jassal
www.reactome.orgMurcia - 3 February 20112
Rationale – Journal informationNature 407(6805):770-6.The Biochemistry of Apoptosis.
“Caspase-8 is the key initiator caspase in the death-receptor pathway. Upon ligand
binding, death receptors such as CD95 (Apo-1/Fas) aggregate and form membrane-
bound signalling complexes (Box 3). These complexes then recruit, through adapter
proteins, several molecules of procaspase-8, resulting in a high local concentration of
zymogen. The induced proximity model posits that under these crowded conditions, the
low intrinsic protease activity of procaspase-8 (ref. 20) is sufficient to allow the various
proenzyme molecules to mutually cleave and activate each other (Box 2). A similar
mechanism of action has been proposed to mediate the activation of several other
caspases, including caspase-2 and the nematode caspase CED-3 (ref. 21).”
How can I access the pathway described here and maybe reuse it?
www.reactome.orgMurcia - 3 February 20113
Nature. 2000 Oct 12;407(6805):770-6.The biochemistry of apoptosis.
Rationale - FiguresA picture paints a thousand words…
but….• Just pixels• Omits key details• Assumes• Fact or Hypothesis?
www.reactome.orgMurcia - 3 February 20114
Reactome is…
Free, online, open-source curateddatabase of pathways and reactions in human biology
Authored by expert biologists, maintained byReactome editorial staff (curators)
Mapped to cellular compartment
www.reactome.orgMurcia - 3 February 20115
Extensively cross-referenced
Tools for data analysis – Pathway Analysis, Expression Overlay, Species Comparison, Biomart…
Used to infer orthologous events in 20 non-human species
Reactome is…
www.reactome.orgMurcia - 3 February 20116
Theory - Reactions
Pathway steps = the “units” of Reactome
= events in biology
TRANSPORTCLASSIC
BIOCHEMICAL
BINDING
DISSOCIATION
DEGRADATION
PHOSPHORYLATION
DEPHOSPHORYLATION
www.reactome.orgMurcia - 3 February 20117
Reaction Example 1: Enzymatic
www.reactome.orgMurcia - 3 February 20118
Reaction Example 2: Transport
REACT_945.4
Transport of Ca++ from platelet dense tubular system to cytoplasm
www.reactome.orgMurcia - 3 February 20119
Other Reaction Types
Binding
Dimerization
Phosphorylation
www.reactome.orgMurcia - 3 February 201110
Reactions Connect into Pathways
OUTPUTINPUT
CATALYST
OUTPUTINPUT
CATALYST
INPUT OUTPUT
CATALYS
T
www.reactome.orgMurcia - 3 February 201111
Human pathway
PMID:5555 PMID:4444
mouse
cow
Direct evidence
Indirect evidencePMID:8976
PMID:1234
Evidence Tracking – Inferred Reactions
www.reactome.orgMurcia - 3 February 201112
Data Expansion - Link-outs From Reactome
• GO • Molecular Function• Compartment• Biological process
• KEGG, ChEBI – small molecules• UniProt – proteins• Sequence dbs – Ensembl, OMIM, Entrez Gene,
RefSeq, HapMap, UCSC, KEGG Gene• PubMed references – literature evidence for events
www.reactome.orgMurcia - 3 February 201113
Species Selection
www.reactome.orgMurcia - 3 February 201114
Data Expansion – Projecting to Other Species
A + ATP A + ADP-PB
Human
A + ATP A + ADP-P
BMouse
BA
Drosophila
Reaction notinferred
No orthologue - Protein not inferred
+ ATP
www.reactome.orgMurcia - 3 February 201115
Exportable Protein-Protein Interactions
Inferred from complexes and reactions
Interactions between proteins in the same complex, reaction, or adjoining reaction
Lists available from Downloads
See Readme document for more details
www.reactome.orgMurcia - 3 February 201116
Coverage – Current TOC
And many more...
www.reactome.orgMurcia - 3 February 201117
Planned Coverage – Editorial Calendar
www.reactome.orgMurcia - 3 February 201118
Reactome Tools
Interactive Pathway Browser
Pathway Mapping and Over-representation
Expression overlay onto pathways
Molecular Interaction overlay
Biomart
www.reactome.orgMurcia - 3 February 201119
Arabidopsis Reactome
http://www.arabidopsisreactome.org/
www.reactome.orgMurcia - 3 February 201120
Tutorial
www.reactome.orgMurcia - 3 February 201121
Front Page http://www.reactome.org
Main text
Navigation bar
www.reactome.orgMurcia - 3 February 201122
Exercise 1
www.reactome.orgMurcia - 3 February 201123
The Pathway BrowserSpecies selector
Search & Analyze barSidebar
Pathway Diagram Panel
Details Panel (hidden)Thumbnail
www.reactome.orgMurcia - 3 February 201124
Pathways tab – pathway hierarchy
PathwayReactionBlack-box
www.reactome.orgMurcia - 3 February 201125
Exercise 2
From the homepage, search for ‘Notch signaling’.Click on the top pathway hit. This will open it in the Pathway Browser.Ignoring the diagram for now, look at the Pathways tab on the left.
1.How many sub-pathways does this pathway have?2.How many reactions are in the first of these sub-pathways?3.What reaction follows Notch 2 precursor transport to Golgi?Hint: If it’s not visible, open the Details pane at the bottom of the
page by clicking on the blue triangle.
www.reactome.orgMurcia - 3 February 201126
The Pathway Browser - Pathway Diagrams
Boxes are proteins, sets or complexes. Ovals are small molecules.Green boxes are proteins or sets, blue are complexes.
Catalyst
Input
Outputs
Compartment
Reaction node
Transition Binding Dissociation Omitted Uncertain
Regulation+ve -ve
www.reactome.orgMurcia - 3 February 201127
Exercise 3
From the Homepage, search for the pathway ‘Effects of PIP2 hydrolysis’ and open it in the Pathway Browser.
1. What symbol represents the reaction for ‘Binding of IP3 to the IP3 receptor’?
2. 2. What symbol represents the reaction ‘Transport of Ca++ from platelet dense tubular system to cytoplasm’? What subtype of reaction is this?
3. 3. What is the catalyst (descriptive name) for ‘2-AG hydrolysis to arachidonate by MAGL’? Can you find its UniProt ID and name the two outputs of this reaction?
www.reactome.orgMurcia - 3 February 201128
Navigating in the Pathway Browser I
Click here
Highlights
Details here
Click here to open pathway diagram...
Home and Analyze buttonsSearch
www.reactome.orgMurcia - 3 February 201129
Highlights
Details here
Navigating in the Pathway Browser II
Click here
Zoom
www.reactome.orgMurcia - 3 February 201130
Exercise 4
www.reactome.orgMurcia - 3 February 201131
The Details Panel
www.reactome.orgMurcia - 3 February 201132
Exercise 5
www.reactome.orgMurcia - 3 February 201133
Pathway Analysis
www.reactome.orgMurcia - 3 February 201134
Pathway Analysis – Overrepresentation
‘Top-level’
Reveal next level
P-val
www.reactome.orgMurcia - 3 February 201135
Exercise 6
Check this!
1. What is the most significantly over-represented top-level pathway for this dataset?
2. How many genes are in this pathway, and how many were represented in the dataset?
3. Why is the top-level pathway Chromosome Maintenance higher in the list than Signalling by Wnt when the latter has a more significant probability score? (Hint – use the Open All button)
4. Can you interpret these results in terms of the underlying biology? (Hint: good luck, there are many correct answers!)
www.reactome.orgMurcia - 3 February 201136
Species Comparison I
www.reactome.orgMurcia - 3 February 201137
Species Comparison II
Yellow = human/rat Blue = human onlyGrey = not relevantBlack = Complex
www.reactome.orgMurcia - 3 February 201138
Exercise 7
www.reactome.orgMurcia - 3 February 201139
Expression Analysis I
www.reactome.orgMurcia - 3 February 201140
Expression Analysis II ‘Hot’ = high‘Cold’ = low
Step throughData columns
www.reactome.orgMurcia - 3 February 201141
Exercise 8
www.reactome.orgMurcia - 3 February 201142
Molecular Interaction Overlay
www.reactome.orgMurcia - 3 February 201143
Exercise 9
Open the pathway diagram for Netrin-1 Signaling.
1.Find the protein FADK1 (top centre of the cytosol). Right click on it and select Display Interactors. How many are there? 2.How many times has the interaction between FADK1 and Tumor endothelial marker 6 been documented? Hint: This detail is not in Reactome.3.Find the protein SRC (to the left of FADK1). Display interactors for this protein. How many are there? Can you get a list of them?4.Display interactors for UNC5B (bottom left of the cytosol). What happens and why?5.What is the easiest way to remove interactors?
www.reactome.orgMurcia - 3 February 201144
BioMart – selecting your dataset
www.reactome.orgMurcia - 3 February 201145
BioMart – filters
www.reactome.orgMurcia - 3 February 201146
BioMart – attributes
Check to get
attribute
www.reactome.orgMurcia - 3 February 201147
BioMart – results
www.reactome.orgMurcia - 3 February 201148
Exercise 10
www.reactome.orgMurcia - 3 February 201149
The End