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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

酿酒酵母高迁移率族B（HMGB）蛋白HM 01与rRNA基因座和许多核糖体蛋白基因（RPGs）的启动子相关，本文利用Sos募集系统证明HM 01与TFIID的组成部分TBP和TAF 1的N-末端结构域（TAND）相互作用。生化研究表明，HM 01与TFIID共纯化，并直接与TBP相互作用，但不与TAND相互作用。HM 01（Deltahmol）的缺失导致严重的冷敏感性生长缺陷，并降低了一些TAND依赖基因的转录。Deltahmol还以启动子特异性方式影响一些RPG启动子处的TFIID占用。有趣的是，HM 01的过表达延迟了缺乏TAND的tafl突变体（taf 1DeltaTAND）的菌落形成，但不是野生型菌株，表明HM 01和TAND之间的功能联系。此外，Deltahmol在一些spt 15（TBP）和toal（TFIIA）突变体中表现出合成性生长缺陷，而它拯救了一些类似的生长缺陷。 ...更多信息 a7（TFIIB）突变体。重要的是，Deltahmol导致RPS 5、RPS 16 A、RPL 23 B、RPL 27 B和RPL 32的转录起始位点向上游移动，但不导致RPS 31、RPL 10、TEF 2和ADH 1的转录起始位点向上游移动，表明HM 01可能通过与TFIID的相互作用参与II类基因亚组的起始位点选择。和RAP 1在RPG转录中的表达。结果显示，RPG启动子（总共138个）可以基于启动子处的HMO 1丰度和FHL 1和/或RAP 1与启动子结合的HMO 1依赖性而被分为几个不同的组。FHL 1以HMO 1依赖性方式与大多数HMO 1富集和转录HMO 1依赖性RPG启动子结合，而以HMO 1非依赖性方式与HMO 1限制性RPG启动子结合，无论它们是否以HMO 1依赖性方式转录。重要的是，这些功能特性由启动子序列决定。少