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Saccharomyces cerevisiae HMO1 interacts with TFIID and participates in start site selection by RNA polymerase II

机译:酿酒酵母HMO1与TFIID相互作用并参与RNA聚合酶II的起始位点选择

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摘要

Saccharomyces cerevisiae HMO1, 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 HMO1 interacts with TBP and the N-terminal domain (TAND) of TAF1, which are integral components of TFIID. Biochemical studies revealed that HMO1 copurifies with TFIID and directly interacts with TBP but not with TAND. Deletion of HMO1 (Δhmo1) causes a severe cold-sensitive growth defect and decreases transcription of some TAND-dependent genes. Δhmo1 also affects TFIID occupancy at some RPG promoters in a promoter-specific manner. Interestingly, over-expression of HMO1 delays colony formation of taf1 mutants lacking TAND (taf1ΔTAND), but not of the wild-type strain, indicating a functional link between HMO1 and TAND. Furthermore, Δhmo1 exhibits synthetic growth defects in some spt15 (TBP) and toa1 (TFIIA) mutants while it rescues growth defects of some sua7 (TFIIB) mutants. Importantly, Δhmo1 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 HMO1 may participate in start site selection of a subset of class II genes presumably via its interaction with TFIID.
机译:啤酒酵母HMO1是高迁移率B组(HMGB)蛋白,与rRNA基因座以及许多核糖体蛋白基因(RPG)的启动子相关。在这里,Sos募集系统用于显示HMO1与TBI和TAF1的N末端域(TAND)相互作用,它们是TFIID的组成部分。生化研究表明,HMO1与TFIID共纯化,并与TBP直接相互作用,但与TAND不相互作用。 HMO1(Δhmo1)的删除会导致严重的冷敏感生长缺陷,并减少某些TAND依赖基因的转录。 Δhmo1还以启动子特异性方式影响某些RPG启动子的TFIID占用。有趣的是,HMO1的过表达延迟了缺乏TAND的taf1突变体(taf1ΔTAND)的集落形成,但没有野生型菌株的集落形成,表明HMO1和TAND之间存在功能联系。此外,Δhmo1在某些spt15(TBP)和toa1(TFIIA)突变体中表现出合成生长缺陷,而它可以挽救某些sua7(TFIIB)突变体的生长缺陷。重要的是,Δhmo1会导致RPS5,RPS16A,RPL23B,RPL27B和RPL32的转录起始位点向上游偏移,但不会导致 RPS31 RPL10 TEF2 ADH1 ,表明HMO1可能通过与TFIID的相互作用参与了II类基因子集的起始位点选择。

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