A TATA-binding protein mutant defective for TFIID complex formation in vivo.
Using an intragenic complementation screen, we have identified a temperature-sensitive TATA-binding protein (TBP) mutant (K151L, K156Y) that is defective for interaction with certain yeast TBP-associated factors (TAFs) at the restrictive temperature. The K151L,K156Y mutant appears to be functional for RNA polymerase I (Pol I) and Pol III transcription, and it is capable of supporting Gal4-activated and Gcn4-activated transcription by Pol II. However, transcription from certain TATA-containing and TATA-less Pol II promoters is reduced at the restrictive temperature. Immunoprecipitation analysis of extracts prepared after culturing cells at the restrictive temperature for 1 h indicates that the K151L,K156Y derivative is severely compromised in its ability to interact with TAF130, TAF90, TAF68/61, and TAF25 while remaining functional for interaction with TAF60 and TAF30. Thus, a TBP mutant that is compromised in its ability to form TFIID can support the response to Gcn4 but is defective for transcription from specific promoters in vivo.
D000640:Amitrole / Q000378:metabolism; D004257:DNA Polymerase II / Q000502:physiology; D004258:DNA Polymerase III / Q000502:physiology; D004791:Enzyme Inhibitors / Q000378:metabolism; D015151:Immunoblotting; D008958:Models, Molecular; D016296:Mutagenesis; D010641:Phenotype; D011233:Precipitin Tests; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016385:TATA Box / Q000502:physiology*; D013696:Temperature; D035362:Transcription Factor TFIID; D019473:Transcription Factors, TFII / Q000502:physiology*; D014158:Transcription, Genetic