title: Identification of a novel TATA element-binding protein binding region at the N terminus of the Saccharomyces cerevisiae TAF1 protein.
abstract: TFIID, a multiprotein complex composed of TATA element-binding protein (TBP) and 14 TBP-associated factors (TAFs), can directly recognize core promoter elements and mediate transcriptional activation. The TAF N-terminal domain (TAND) of TAF1 may play a significant role in these two principal TFIID functions by regulating the access of TBP to the TATA element. In yeast, TAND consists of two subdomains, TAND1 (10-37 amino acids (aa)) and TAND2 (46-71 aa), which interact with the concave and convex surfaces of TBP, respectively. Here we demonstrate that another region located on the C-terminal side of TAND2 (82-139 aa) can also bind to TBP and induce transcriptional activation when tethered to DNA as a GAL4 fusion protein. As these properties are the same as those of TAND1, we denoted this sequence as TAND3. Detailed mutational analyses revealed that three blocks of hydrophobic amino acid residues located within TAND3 are required not only for TBP binding and transcriptional activation but also for supporting cell growth and the efficient transcription of a subset of genes. We also show that the surface of TBP recognized by TAND3 is broader than that recognized by TAND1, although these regions overlap partially. Supporting these observations is that TAND1 can be at least partly functionally substituted by TAND3.
authors.affiliation: Division of Molecular and Cellular Biology, Graduate School of Integrated Science, Yokohama City University, Yokohama 230-0045, USA.
authors.forename: Shinya
authors.initials: S
authors.lastname: Takahata
authors.forename: Hidei
authors.initials: H
authors.lastname: Ryu
authors.forename: Kazushige
authors.initials: K
authors.lastname: Ohtsuki
authors.forename: Koji
authors.initials: K
authors.lastname: Kasahara
authors.forename: Masashi
authors.initials: M
authors.lastname: Kawaichi
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Kokubo
mesh_terms: D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins; D004330:Drosophila; D005982:Glutathione Transferase / Q000378:metabolism; D015151:Immunoblotting; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism; D008957:Models, Genetic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D035362:Transcription Factor TFIID / Q000737:chemistry; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C409889:ARN2 protein, S cerevisiae; D002843:Chromatin; D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D004247:DNA; D005982:Glutathione Transferase; D001616:beta-Galactosidase; D000409:Alanine