Studies on the regulatory mechanisms of TFIID-mediated eukaryotic transcription

TFIID介导的真核转录调控机制研究

基本信息

批准号：
18370072
负责人：
KOKUBO Tetsuro
金额：
$ 9.09万
依托单位：
Yokohama City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18370072/
关键词：
transcriptional regulation transcription factor general transcription factor gene expression budding yeast TFIID TAF TATA box binding protein

项目摘要

Saccharomyces cerevisiae HM01, a high mobility group B (HMGB) protein, associates with the rRNA locus and with the promoters of many ribosomal protein genes (RPGs) .Here, the Sos recruitment system was used to show that HM01 interacts with TBP and the N-terminal domain (TAND) of TAF1, which are integral components of TFIID. Biochemical studies revealed that HM01 copurifies with TFIID and directly interacts with TBP but not with TAND. Deletion of HM01 (Deltahmol) causes a severe cold-sensitive growth defect and decreases transcription of some TAND-dependent genes. Deltahmol also affects TFIID occupancy at some RPG promoters in a promoter-specific manner. Interestingly, over-expression of HM01 delays colony formation of tafl mutants lacking TAND (taf1DeltaTAND), but not of the wild-type strain, indicating a functional link between HM01 and TAND. Furthermore, Deltahmol exhibits synthetic growth defects in some spt15 (TBP) and toal (TFIIA) mutants while it rescues growth defects of some su … More a7 (TFIIB) mutants. Importantly, Deltahmol causes an upstream shift in transcriptional start sites of RPS5, RPS16A, RPL23B, RPL27B and RPL32, but not of RPS31, RPL10, TEF2 and ADH1, indicating that HM01 may participate in start site selection of a subset of class II genes presumably via its interaction with TFIID.This study also uses genome-wide chromatin immunoprecipitation to study the roles of HMO1, FHL1, and RAP1 in RPG transcription. The results show that RPG promoters(138 in total) can be classified into several distinct groups based on HM01 abundance at the promoter and the HM01 dependence of FHL1 and/or RAP1 binding to the promoter. FHL1 binds to most of the HMO1-enriched and transcriptionally HMO1-dependent RPG promoters in an HMO1-dependent manner, whereas it binds to HMO1-limited RPG promoters in an HMO1-independent manner, irrespective of whether they are transcribed in an HMO1-dependent manner. Importantly, these functional properties are determined by the promoter sequence. Less

酿酒酵母 HM01 是一种高迁移率 B 族 (HMGB) 蛋白，与 rRNA 基因座和许多核糖体蛋白基因 (RPG) 的启动子相关。这里，使用 Sos 招募系统表明 HM01 与 TBP 和 N 相互作用TAF1 的末端结构域 (TAND) 是 TFIID 的组成部分生化研究表明 HM01 与 TFIID 共纯化。 HM01 (Deltahmol) 的缺失会导致严重的冷敏感生长缺陷，并减少一些 TAND 依赖性基因的转录，并且会以启动子特异性的方式影响一些 RPG 启动子上的 TFIID 占据。 HM01 的阴性、过度表达会延迟缺乏 TAND 的 tafl 突变体 (taf1DeltaTAND) 的集落形成，但不会延迟野生型菌株的集落形成，表明存在功能性联系此外，Deltahmol 在一些 spt15 (TBP) 和 toal (TFIIA) 突变体中表现出合成生长缺陷，同时它可以挽救一些 su … 更多 a7 (TFIIB) 突变体的生长缺陷。 RPS5、RPS16A、RPL23B、RPL27B 和 RPL32 位点，但不包括 RPS31、RPL10、 TEF2 和 ADH1，表明 HM01 可能通过与 TFIID 相互作用参与 II 类基因子集的起始位点选择。本研究还使用全基因组染色质免疫沉淀来研究 HMO1、FHL1 和 RAP1 在 RPG 转录中的作用结果表明，根据启动子处的 HM01 丰度和 FHL1 的 HM01 依赖性，RPG 启动子（总共 138 个）可以分为几个不同的组。和/或 RAP1 与启动子的结合。重要的是，这些功能特性是由启动子序列决定的。