The RNA polymerase II general transcription factor TFIID is a complex containing the TATA-binding protein (TBP) and associated factors (TAFs). We have used a mutant allele of the gene encoding yeast TAF(II)68/61p to analyze its function in vivo. We provide biochemical and genetic evidence that the C-terminal alpha-helix of TAF(II)68/61p is required for its direct interaction with TBP, the stable incorporation of TBP into the TFIID complex, the integrity of the TFIID complex, and the transcription of most genes in vivo. This is the first evidence that a yeast TAF(II) other than TAF(II)145/130 interacts with TBP, and the implications of this on the interpretation of data obtained studying TAF(II) mutants in vivo are discussed. We have identified a high copy suppressor of the TAF68/61 mutation, TSG2, that has sequence similarity to a region of the SAGA subunit Ada1. We demonstrate that it directly interacts with TAF(II)68/61p in vitro, is a component of TFIID, is required for the stability of the complex in vivo, and is necessary for the transcription of many yeast genes. On the basis of these functions, we propose that Tsg2/TAF(II)48p is the histone 2A-like dimerization partner for the histone 2B-like TAF(II)68/61p in the yeast TFIID complex.
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| Gene/Complex | Qualifier | Gene Ontology Term | Annotation Extension | Evidence | Source | Assigned On |
|---|---|---|---|---|---|---|
| TAF12 | involved in | transcription by RNA polymerase II | IMP | SGD | 2013-08-07 | |
| TAF12 | enables | TBP-class protein binding | IDA | SGD | 2013-08-07 |
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| Gene | Phenotype | Experiment Type | Mutant Information | Strain Background | Chemical | Details |
|---|---|---|---|---|---|---|
| TAF12 | heat sensitivity: increased | classical genetics | conditional Allele: taf12-(taf68-9) W486stop | Other | Temperature: elevated temperature, 37 °C |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
|---|
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| Interactor | Interactor | Assay | Annotation | Action | Modification | |
|---|---|---|---|---|---|---|
| SPT15 | TAF1 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| SPT15 | TAF12 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| SPT15 | TAF4 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| SPT15 | TAF1 | Reconstituted Complex | manually curated | Hit-Bait | No Modification | |
| SPT15 | TAF12 | Reconstituted Complex | manually curated | Hit-Bait | No Modification | |
| SPT7 | TAF10 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| SPT7 | TAF12 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| SPT7 | TAF5 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| TAF1 | TAF4 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| TAF12 | TAF1 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification |