Functional characterization of TFIID subunits (TAFs) in Saccharomyces cerevisiae

酿酒酵母 TFIID 亚基 (TAF) 的功能表征

• 批准号: 11680680
• 负责人: KOKUBO Tetsuro
• 金额: $ 2.37万
• 依托单位: NARA INSTITUTE OF SCIENCE AND TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680680/
• 关键词: transcription; tanscriptional regulation; TFIID; TAF; general transcription factor; yeast; basal transcription; core promoter

The general transcription factor TFIID, which is composed of a TATA-binding protein (TBP) and a set of TBP-associated factors (TAFs), has been shown to be involved in both core promoter recognition and the transcriptional activation of eukaryotic genes. We isolated TAF145 temperature-sensitive mutants in yeast, in which transcription of the TUB2 gene is impaired at restrictive temperatures due to a defect in core promoter. We also showed in these mutants that the transcription of the RPS5 gene is impaired, mostly due to a defect in transcriptional activation rather than to a defect in core promoter recognition, although the latter is slightly affected as well. Surprisingly, the RPS5 core promoter can be activated by various activation domains fused to a GAL4 DNA binding domain (e.g. GAL4-RAP1, -VP16C, -EBNA2, and -GCN4), but not by the original upstream activating sequence (UAS) of the RPS5 gene. In addition, a heterologous CYC1 core promoter can be activated by RPS5-UAS at normal levels even in these mutants. These observations indicate that a distinct combination of core promoters and activators may exploit alternative activation pathways which vary in their requirement for TAF 145 function. In addition, a particular function of TAF 145 that is deleted in our mutants appears to be involved in both core promoter recognition and transcriptional activation

一般转录因子TFIID由一个tata结合蛋白（TBP）和一组TBP相关因子（TAFs）组成，已被证明参与核心启动子识别和真核基因的转录激活。我们从酵母中分离出TAF145温度敏感突变体，其中TUB2基因的转录由于核心启动子的缺陷而在限制性温度下受损。我们还发现，在这些突变体中，RPS5基因的转录受损，主要是由于转录激活缺陷，而不是由于核心启动子识别缺陷，尽管后者也受到轻微影响。令人惊讶的是，RPS5核心启动子可以被融合到GAL4 DNA结合域的各种激活域（例如GAL4- rap1， -VP16C， -EBNA2和-GCN4）激活，但不能被RPS5基因的原始上游激活序列（UAS）激活。此外，即使在这些突变体中，异源CYC1核心启动子也可以被正常水平的RPS5-UAS激活。这些观察结果表明，核心启动子和激活子的不同组合可能利用不同的TAF 145功能需求的替代激活途径。此外，在我们的突变体中缺失的TAF 145的特定功能似乎涉及核心启动子识别和转录激活

项目成果

Kobayashi, A., Miyake, T., Ohyama, Y., Kawaichi, M., and Kokubo, T.: "Mutations in the TATA binding protein, affecting transcriptional activation, show synthetic lethality with the TAF145 gene lacking the TAF N-terminal domain in Saccharomyces cerevisiae.

Kobayashi, A.、Miyake, T.、Ohyama, Y.、Kawaichi, M. 和 Kokubo, T.：“影响转录激活的 TATA 结合蛋白突变，对缺乏 TAF N- 的 TAF145 基因表现出合成致死性”

C.R.Lim,Y.Kimata,H.Ohdata,T.Kobubo,N.Kikuchi,T.Horigome,K.Kohno: "The Saccharomyces cerevisiae RuvB-like protein, tih2p.is required for cell cycle progression and RNA polymerase II-directed transcription"Journal of Biological Chemistry. 275・29. 22409-2241

C.R.Lim、Y.Kimata、H.Ohdata、T.Kobubo、N.Kikuchi、T.Horigome、K.Kohno：“酿酒酵母 RuvB 样蛋白 tih2p. 是细胞周期进展和 RNA 聚合酶 II 指导所必需的转录“生物化学杂志.275・29.22409-2241

T.Kotani,K-i.Banno,M.Ikura,A.G.Hinnebusch,Y.Nakatani,M.Kawaichi,T.Kokubo: "A role of transcriptional activators as anti-repressors for the auto-inhibitory activity of TATA box binding of TFIID"Proc.Natl.Acad.Sci.USA. 97・13. 7178-7183 (2000)

T. Kotani、K-i. Banno、M. Ikura、A. G. Hinnebusch、Y. Nakatani、M. Kawaichi、T. Kokubo：“转录激活剂作为 TFIID 的 TATA 盒结合的自动抑制活性的抗阻遏物的作用”美国国家科学院院刊 97・13。

Kotani, T., Banno, K-i., Ikura, M., Hinnebusch, A.G., Nakatani, Y., Kawaichi, M., _and Kokubo, T.: "A role of transcriptional activators as anti-repressors for the auto-inhibitory activity of TATA box binding of TFIID"Proc.Natl.Acad.Sci.USA. 97-13. 7178-7

Kotani, T.、Banno, K-i.、Ikura, M.、Hinnebusch, A.G.、Nakatani, Y.、Kawaichi, M., _and Kokubo, T.：“转录激活剂作为自身抑制的抗阻遏物的作用

A.Kobayashi,T.Miyake,Y.Ohyama,M.Kawaichi,T.Kokubo: "Mutations in the TATA binding protein, affecting transcriptional activation, show synthetic lethality with the TAF145 gene lacking the TAFN-terminal domain in Saccharomyces cerevisiae"Journal of Biologic

A.Kobayashi、T.Miyake、Y.Ohyama、M.Kawaichi、T.Kokubo：“TATA 结合蛋白的突变影响转录激活，在酿酒酵母中缺乏 TAFN 末端结构域的 TAF145 基因显示出合成致死性”杂志