Annotation Conf. Call 2016-01-12: Difference between revisions
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*Reminder that ontology requests for 'response to x' terms that are non-physiological are inappropriate. | *Reminder that ontology requests for 'response to x' terms that are non-physiological are inappropriate. | ||
*Please see github ticket: [https://github.com/geneontology/go-ontology/issues/12228 Response to compounds used as assays #12228] | *Please see github ticket: [https://github.com/geneontology/go-ontology/issues/12228 Response to compounds used as assays #12228] | ||
==Annotation from Abstracts Only== | |||
*From ontology meeting minutes: We thought that annotations from abstracts only (where the full text of the paper is in non-English language) were discouraged. Looks like this may not be the case for some MODs. But then, how can people verify, a posteriori, correctness of annotations? | |||
*We would like to review what MODs do wrt annotating abstracts to make sure we are all on the same page with this. | |||
*Are there any circumstances where we think this is warranted? | |||
==Deprecated Annotation Extensions== | ==Deprecated Annotation Extensions== | ||
*Rachael and Ruth have put together documentation on the deprecated extensions along with [http://wiki.geneontology.org/index.php?title=Guidance_for_updating_deprecated_Annotation_Extension_Relations recommendations] for revision. | *Rachael and Ruth have put together documentation on the deprecated extensions along with [http://wiki.geneontology.org/index.php?title=Guidance_for_updating_deprecated_Annotation_Extension_Relations recommendations] for revision. |
Revision as of 15:24, 9 January 2016
Agenda
New Year's Welcome
- Plans for upcoming year
- Annotation Consistency Exercises
- Will continue monthly exercises for 2016
- This year, we will add discussion of LEGO representation to the calls, and compare conventional GO annotations to LEGO representations.
- AI: Confirm rota schedule.
- Annotation Consistency Exercises
Ontology Updates
- Reminder that ontology requests for 'response to x' terms that are non-physiological are inappropriate.
- Please see github ticket: Response to compounds used as assays #12228
Annotation from Abstracts Only
- From ontology meeting minutes: We thought that annotations from abstracts only (where the full text of the paper is in non-English language) were discouraged. Looks like this may not be the case for some MODs. But then, how can people verify, a posteriori, correctness of annotations?
- We would like to review what MODs do wrt annotating abstracts to make sure we are all on the same page with this.
- Are there any circumstances where we think this is warranted?
Deprecated Annotation Extensions
- Rachael and Ruth have put together documentation on the deprecated extensions along with recommendations for revision.
Please document here examples of where you have questions about re-annotation of annotations with deprecated extensions.
dependent_on
Example 1
- Co-transfection experiment that used dependent_on relation to capture requirement of both proteins to effect a GPCR signaling pathway
- Recommendation: make direct annotations to each gene product to avoid 'reverse' annotations and use contributes_to qualifier
- Two chemoreceptors mediate developmental effects of dauer pheromone in C. elegans.
- To investigate whether expression of SRBC-64/66 was sufficient to confer pheromone responses in a heterologous context, we expressed one or both receptors in HEK293 cells and monitored intracellular second messenger levels upon the addition of ascarosides. Co-expression of SRBC-64 and -66, but not each receptor alone, enhanced forskolin-mediated increases in cAMP levels (Fig. 4A, B), and this increase was significantly inhibited upon addition of C6 and C9 at different concentrations (Fig. 4A-C).
- Old annotation (one of four annotations made):
Gene product: SRBC-64 (UniProtKB:G5ECV4) GO ID: G-protein coupled receptor activity (GO:0004930) GO evidence: IDA Extension: dependent_on SRBC-66 (UniProtKB:O61938)
- Possible new annotation:
Gene product: SRBC-64 (UniProtKB:G5ECV4) GO qualifier: contributes_to GO ID: G-protein coupled receptor activity (GO:0004930) GO evidence: IDA
In this case, I will lose the information that the activity is only seen in the presence of SRBC-66.
- Possible new annotation:
Gene product: SRBC-64 (UniProtKB:G5ECV4) GO qualifier: contributes_to GO ID: G-protein coupled receptor activity (GO:0004930) GO evidence: IGI With: SRBC-66 (UniProtKB:O61938)
Should this instead be an IGI annotation?
Example 2
- Co-transfection experiment demonstrating need for a tethering molecule to facilitate enzymatic activity
- Recommendation: make direct annotations to avoid 'reverse' annotations and use contributes_to qualifier
- LIN-28 and the poly(U) polymerase PUP-2 regulate let-7 microRNA processing in Caenorhabditis elegans.
- PUP-2 was previously shown to polyuridylate an artificially tethered RNA in Xenopus oocytes but was inactive without tethering. Therefore, we tested whether LIN-28 might be able to recruit PUP-2 to mediate pre–let-7 uridylation (Fig. 3c). We incubated anti-Flag immunoprecipitates from cell extracts expressing HA-Flag–PUP-2 and/or SBP–LIN-28 with radiolabeled pre–let-7 and radiolabeled UTP. HA-Flag–PUP-2 uridylated pre–let-7 only in the presence of SBP–LIN-28 (Fig. 3c). We also confirmed LIN-28–dependent uridylation of pre–let-7 by PUP-2 in vitro (Supplementary Fig. 6b).
- Old annotation:
Gene product: PUP-2 (UniProtKB:Q09408) GO ID: RNA uridylyltransferase activity (GO:0050265) GO evidence: IDA Extension: dependent_on LIN-28 (UniProtKB:P92186)
- Possible new annotations:
Gene product: PUP-2 (UniProtKB:Q09408) GO ID: RNA uridylyltransferase activity (GO:0050265) GO evidence: IDA
Gene product: LIN-28 (UniProtKB:P92186) GO qualifier: contributes_to GO ID: RNA uridylyltransferase activity (GO:0050265) GO evidence: IDA
Gene product: PUP-2 (UniProtKB:Q09408) GO ID: protein complex (GO:0043234) GO evidence: IDA
Gene product: LIN-28 (UniProtKB:P92186) GO ID: protein complex (GO:0043234) GO evidence: IDA
- OR
Gene product: PUP-2 (UniProtKB:Q09408) GO ID: RNA uridylyltransferase activity (GO:0050265) GO evidence: IGI With: LIN-28 (UniProtKB:P92186)
Gene product: LIN-28 (UniProtKB:P92186) GO qualifier: contributes_to GO ID: RNA uridylyltransferase activity (GO:0050265) GO evidence: IGI With: PUP-2 (UniProtKB:Q09408)
Example 3
- Enzymatic activity requires presence of another protein that acts as an upstream activator
- Recommendation: make direct annotations to avoid 'reverse' annotations and use contributes_to qualifier
- Since SGK-1, AKT-1, and AKT-2 all contain a highly similar kinase domain, we tested if they are able to phosphorylate their candidate substrate DAF-16 in vitro. For this purpose, we constructed transgenic wild-type and pdk-1(sa680) mutant strains expressing gfp-tagged akt-1, akt-2, or sgk-1. The fusion proteins were immunopurified from worm extracts and used for kinase assays with purified DAF-16. Both AKT-1, AKT-2, and SGK-1 purified from a wild-type background phosphorylated DAF-16 ( Figure 4A). In contrast, all three kinases purified from a pdk-1 mutant background were not able to phosphorylate DAF-16 (Figure 4A). This demonstrates that functional PDK-1 is required to activate AKT-1, AKT-2, and SGK-1 in vivo.
- In pull-down experiments, we found that both in vitro-translated PDK-1 and DAF-16 interact with bacterially expressed, GST-tagged SGK-1, AKT-1, and AKT-2.
- Old annotation (one example):
Gene product: AKT-1 (UniProtKB:Q17941) GO ID: protein kinase activity (GO:0004672) GO evidence: IDA Extension: has_direct_input DAF-16 (UniProtKB:O16850),dependent_on PDK-1 (UniProtKB:Q9Y1J3)
- New annotations:
Gene product: AKT-1 (UniProtKB:Q17941) GO ID: protein kinase activity (GO:0004672) GO evidence: IDA Extension: has_direct_input DAF-16 (UniProtKB:O16850)
Gene product: PDK-1 (UniProtKB:Q9Y1J3) GO ID: protein kinase activator activity (GO:0030295) GO evidence: IMP Extension: has_direct_input AKT-1 (UniProtKB:Q17941)
Example 4
- Protein interaction that only occurs in the presence of a specific DNA sequence
- Recommendation: use occurs_at
- The Caenorhabditis elegans JNK signaling pathway activates expression of stress response genes by derepressing the Fos/HDAC repressor complex.
- Next we asked whether FOS-1 could interact with HDA-1. T7-FOS-1 and FLAG-HDA-1 were co-expressed in HEK293 cells. We immunoprecipitated FLAG-HDA-1 with anti-FLAG antibodies, and probed for the T7-FOS-1 on a Western blot with anti-T7 antibodies. We failed to detect an association between FOS1- and HDA-1 (Figure 7C, lane 1). However, if we transfected in the Pkreg-1::venus reporter along with T7-FOS-1 and FLAG-HDA-1, we could detect an association between FOS-1 and HDA-1 (Figure 7C, lane 3). Furthermore, removal of the TRE2 site from the Pkreg-1::venus reporter reduced this interaction (Figure 7C, lane 4). These results suggest that HDA-1 and FOS-1 can associate on the kreg-1 promoter.
- Old annotation (one of two reciprocal annotations made):
Gene product: FOS-1 (UniProtKB:G5ECG2) GO ID: histone deacetylase binding (GO:0042826) GO evidence: IPI With/from: HDA-1 (UniProtKB:O17695) Extension: dependent_on RNApol_II_promoter (SO:0000170),dependent_on kreg-1 gene (WB:WBGene00018725)
- New annotation:
Gene product: FOS-1 (UniProtKB:G5ECG2) GO ID: histone deacetylase binding (GO:0042826) GO evidence: IPI With/from: HDA-1 (UniProtKB:O17695) Extension: occurs_at RNApol_II_promoter (SO:0000170) Gene IDs are not currently valid for occurs_at relation, so cannot add the specific gene. Only GO and SO IDs are currently valid.