Study on the mechanism of transcriptional regulation by the general transcription factor TFIID (The Role of WD

通用转录因子TFIID转录调控机制研究(The Role of WD)

• 批准号: 14540569
• 负责人: MITSUZAWA Hiroshi
• 金额: $ 2.24万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14540569/
• 关键词: general transcription factor; TFIID; TAF; WD repeat; fission yeast; transcriptional regulation; SAGA; Gcn5

The general transcription factor TFIID, which consists of TATA-binding protein (TBP) and TBP-associated factors (TAF5), plays a central role in transcription initiation by RNA polymerase II in eukaryotes. This study was carried out to understand the mechanism of transcriptional regulation by TFIID, focusing on the WD repeat-containing TAFs, TAF72 and TAF73, of the fission yeast Schizosaccharomyces pombe.TAF72 was used as bait in a two-hybrid screen to identify its interacting proteins. The screen yielded hitone H4-like TAF with a calculated molecular mass of 50 kDa, which was named TAF50. Coimmunoprecipitation experiments demonstrated that TAF50, as well as TAF72, is a component of the S. pombe TFIID complex. Both TAFs were also present in the SAGA complex, a histone acetyltransferase complex distinct from TFIID.To identify additional TAFs in S. pombe, database searches were performed, revealing that the S. pombe genome contains a set of TAFs homologous to those identified in other organisms.In addition, temperature-sensitive mutants of TAF72 and TAF73 were isolated to elucidate the in vivo function of these TAFs.These results uncover a function of the WD-repeat region of TAF72, that is, mediating interaction between TAFs, and also provide information about the basal transcription machinery of S. pombe. Furthermore, the temperature-sensitive mutants of TAF72 and TAF73 isolated in this study would serve as materials for future work.

通用转录因子TFIID由TATA结合蛋白(TBP)和TBP相关因子(TAF5)组成，在真核生物的RNA聚合酶II的转录启动中起着核心作用。本研究以裂殖酵母裂殖酵母中含有WD重复序列的TAF72和TAF73为研究对象，通过双杂交筛选TAF72作为诱饵，对TFIID的转录调控机制进行了研究。筛选得到HitOne-H4类TAF，计算其分子质量为50 kDa，命名为TAF50。免疫共沉淀实验证明，TAF50和TAF72都是S.pombe TFIID复合体的一个组成部分。这两个TAFs也存在于SAGA复合体中，这是一个有别于TFIID的组蛋白乙酰转移酶复合体。为了鉴定更多的TAFs，我们进行了数据库搜索，发现了一组与其他组织中发现的TAFs同源的TAFs。此外，我们还分离出了TAF72和TAF73的温度敏感突变体，以阐明这些TAFs在体内的功能。这些结果揭示了TAF72的WD-重复区域的一个功能，即介导TAFs之间的相互作用，并提供了关于S.pombe的基本转录机制的信息。此外，本研究分离到的TAF72和TAF73的温敏突变株可作为后续工作的材料。

项目成果

光澤 浩: "分裂酵母を用いた基本転写因子TFIIDの解析"蛋白質 核酸 酵素. 47・14. 1931-1938 (2002)

Hiroshi Mitsuzawa：“使用裂殖酵母分析基本转录因子 TFIID”蛋白质核酸酶 47・14（2002 年）。

Analysis of the general transcription factor TFIID using fission yeast

使用裂殖酵母分析通用转录因子 TFIID

• 发表时间: 2002
• 期刊: Tanpakushitsu Kakusan Koso 47(14)
• 作者: Hiroshi Mitsuzawa;Yukiko Imazawa;Hiroshi Mitsuzawa;Makoto Kimura;Hiroshi Mitsuzawa;Makoto Kimura;Hiroshi Mitsuzawa;光澤 浩;Hiroshi Mitsuzawa;Hiroshi Mitsuzawa
• 通讯作者: Hiroshi Mitsuzawa

Hiroshi Mitsuzawa: "Rpb7 subunit of RNA polymerase II interacts with an RNA-binding protein involved in processing of transcripts"Nucleic Acids Research. 31・16. 4696-4701 (2003)

Hiroshi Mitsuzawa：“RNA 聚合酶 II 的 Rpb7 亚基与参与转录本加工的 RNA 结合蛋白相互作用”，《核酸研究》31・16 (2003)。

RNA polymerases and accessory factors

RNA聚合酶和辅助因子

• 发表时间: 2004
• 期刊: Molecular Biology of Schizosaccharomyces pombe (Springer-Verlag)
• 作者: Hiroshi Mitsuzawa;Yukiko Imazawa;Hiroshi Mitsuzawa;Makoto Kimura
• 通讯作者: Makoto Kimura

分裂酵母を用いた基本転写因子TFIIDの解析

使用裂殖酵母分析基本转录因子 TFIID

• 发表时间: 2002
• 期刊: 蛋白質 核酸 酵素 47・14
• 作者: Hiroshi Mitsuzawa;Yukiko Imazawa;Hiroshi Mitsuzawa;Makoto Kimura;Hiroshi Mitsuzawa;Makoto Kimura;Hiroshi Mitsuzawa;光澤 浩
• 通讯作者: 光澤 浩