CACAO - Penn State Gene Function and Gene Ontology January

What is an annotation? Dictionary.com: a critical or explanatory note or body of notes added to a text For us, it is adding biologically relevant information to a protein record

Function annotation Allows us to – Infer the functions of genes Related by common descent Related by similar expression patterns Related by phylogenetic profiles...

Function annotation Allows us to – Understand the capabilities of organisms genomes – Understand patterns of gene expression In different environments In different tissues In disease states –...

Classic MODel Literature Datasets Curators (rate limiting) Database

Requirements Accurate functional annotation for as many genes as possible A system of assigning function that allows both humans and computers to compare, contrast, analyze, and predict gene function Curators to make and/or check these assignments – For CACAO, we will teach you what biocurators do.

What’s in it for you (besides credit)? – We hope you will learn how we think about gene function gain skills that will help your future career enjoy contributing to a resource used by people all over the world have fun!

CACAO Community Assessment – How well can Community – you (with our coaching) Annotation with – assign gene functions Ontologies – using GO?

GO = Gene Ontology Controlled vocabulary – Everyone uses the same terms – Terms have IDs that computers can understand Relationships between functions

GO 3 aspects (ontologies) for gene products 1. Biological Process 2. Molecular Function 3. Cellular Component Used to make annotations –aka Gene associations –Term + qualifiers + evidence code + reference etc.

Molecular Function activities or “jobs” of a gene product glucose-6-phosphate isomerase activity from GOC figure from GO consortium presentations

Biological Process a commonly recognized series of events cell division Figure from Nature Reviews Microbiology 6, (January 2008)

Cellular Component where a gene product acts

Key elements of a GO annotation Submitted to GO consortium Viewable on GONUTS **Don’t worry - I will cover this again (several times)!

GO Annotation To make an annotation, you need to – Assign GO terms to genes (gene products) At appropriate level of specificity Sometimes with Qualifiers – NOT – Contributes_to – Colocalizes_with – Record the evidence

Record the evidence Where it came from: – Reference (database accession) PMID: Kind of evidence: – Evidence codes IMP: Inferred from Mutant Phenotype IDA: Inferred from Direct Assay …

Community Annotation CACAO - the “Community Annotation” part What I am going to tell you about next is: 1. How to choose proteins to annotate 2. Finding GO terms & navigating a GO term page 3. Finding UniProt accessions 4. Making gene pages on GONUTS & the anatomy of a gene page 5. How and where to add an annotation 6. Where to look for your annotations & other teams’ annotations … (& the challenges!)

GONUTS Community-editable database GO terms Place to annotate “GoPageMaker” –Makes gene pages with minimal info required –“Annotation” table Editable (by YOU!!!) Pulls information/annotations for proteins from a non- editable database called UniProt

Deciding what to annotate 1. randomly 2. topics of interest (ie efflux pump proteins, biofilms) 3. papers you have come across while doing other stuff 4. methods you know or want to learn 5. phenotypes and mutants you are interested in 6. by author 7. by pathway or regulon 8. suggested by another (ie high IEA:manual annotation ratio) 9. current paper mentions another gene product 10. review papers (ie Annual Reviews are excellent sources) EXAMPLE #1: let’s say you have a great paper (PMID:1111) that characterizes the tyrosine kinase activity of your favorite protein (human p53)…

Finding genes/proteins to annotate UniProt - Textbook or class notes Wikipedia or Google Paper you are reading that mentions another gene –Review articles WikiPathways - PubMed - Ask a coach (usually me) GONUTS –what proteins have other teams been annotating?!

Finding papers PubMed - GoogleScholar References in your textbook Wikipedia & Google Papers given out in another class Papers from a lab you are interested in –Undergrad research work? ** only original research papers ** - no review articles, no textbooks, no books, no class notes

Key elements of a GO annotation Submitted to GO consortium Viewable on GONUTS

Part I: Where do you search for GO terms? GONUTS

CHICK - AgBase (Gallus gallus) dictyBase - dictyBase (Dictyostelium discoideum - slime mold) FB - FlyBase (Drosophila melanogaster) HUMAN - Reactome, BHF-UCL MGI - Mouse genome informatics (Mus musculus - house mouse) SGD - Saccharomyces genome database (Saccharomyces cerevisiase - yeast) TAIR - The Arabidopsis Informatics Resource (Arabidopsis thaliana) WB - WormBase (Caenorhabditis elegans) ZFIN - Zebrafish model organism database (Danio rerio)

What do you actually need once you have found the correct term? GO:

Part II: You now have a paper, a protein & you found a suitable GO term… what next? UniProt accession Search (“ Query ”) & find the correct UniProt accession for your protein - Look something like: P012A9

Part III: Where are you going to add your annotations? GONUTS

How do you make a new gene page in GONUTS? Use the UniProt accession to make a page that you will be able to add your own annotation to. GoPageMaker will: 1.Check if the page exists in GONUTS & take you there if it does. 2.Make a page & pull all of the annotations from UniProt into a table that you can edit.

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… Part IV: Where do you add an annotation? Add a row in the table.

Part V: What you must fill in (for every annotation) GO: PMID:1111 IDA: Inferred from direct assay Figure 2a

What you might also have to fill in Not sure? Check the competition guidelines. Ask a coach (Jim, Debby, Adrienne or usually me)!

Where will your annotation now show up? 1.In the “Annotation” table on the gene page you just edited 2.In the table on your user page 3.In the table on your team page 4.As points on the scoreboard 5.If challenged, it will show up in the “Submitted Challenges” table (below the scoreboard)

CACAO is competitive Teams get points for complete annotations –GO term (right level of specificity) –reference –evidence code –identify where in the paper the evidence comes from Teams can take away points from competitors by challenging annotations –finding a problem –suggesting a better alternative

Community Assessment CACAO - the “Community Assessment” part …

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Scoreboard Submitted Challenges Closed Challenges Moving through challenges

Category:Team UCL1