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Volumn 288, Issue 5469, 2000, Pages 1244-1248

TAF-containing and TAF-independent forms of transcriptionally active TBP in vivo

Author keywords

[No Author keywords available]

Indexed keywords

TATA BINDING PROTEIN;

EID: 0034686037     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5469.1244     Document Type: Article
Times cited : (150)

References (38)
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    • note
    • We generated isogenic yeast strains in which the genes encoding these TAFs were replaced by epitope-tagged versions containing three copies of the HA-1 epitope and performed chromatin immunoprecipitation and quantitative analysis (4). For all four TAFs tested, we observed occupancy of various Pol II promoters in strains containing epitope-tagged TAFs but not in control cells containing untagged TAFs. TAFs were not associated with a tRNA gene promoter, which is transcribed by Pol III, or with a centrally located segment of the POL1 structural gene. When compared on the RPL9A promoter, crosslinking efficiencies of the various TAFs were about the same (0.2% to 0.4%). TBP occupancy on the same samples was determined by immunoprecipitation with TBP antibodies.
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    • note
    • For ribosomal protein gene promoters, the TAF/TBP occupancy ratio is defined on nonshocked cells, because heat shock dramatically decreases transcription (24) and TBP occupancy. However, even under heat shock conditions, the residual transcription is associated with a TAF/TBP ratio of 1.
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    • In accord with this suggestion, heat shock causes a Gcn5-dependent increase in histone H3 acetylation at a variety of inducible promoters (E. vomBaur, J. Deckert, K. Struhl, unpublished observations).
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    • note
    • Theoretically, all promoters that interact exclusively with TFIID should have the maximal TAF/TBP occupancy ratio. The ratio of 1.0 used here is arbitrarily defined, and it represents the maximal TAF/TBP occupancy ratio observed in a variety of promoters. It seems likely that promoters with a ratio of 1.0 interact exclusively with TFIID, although we cannot exclude the possibility that other promoters have higher TAF/TBP occupancy ratios.
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    • note
    • Supported by grants from the Human Frontiers Science Program (L.K. and M.M), the Damon Runyon-Walter Winchell Cancer Research Foundation (P.K.), and NIH grant GM30186 (K.S.). We thank Z. Moqtaderi for fruitful discussions and comments on the manuscript and D. Gewirth for TFIIA and TFIIB used to make antibodies.


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