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TAF11与TFIIA相互作用的图谱绘制及功能表征

Mapping and functional characterization of the TAF11 interaction with TFIIA.

作者信息

Robinson M M, Yatherajam G, Ranallo R T, Bric A, Paule M R, Stargell L A

机构信息

Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523-1870, USA.

出版信息

Mol Cell Biol. 2005 Feb;25(3):945-57. doi: 10.1128/MCB.25.3.945-957.2005.

Abstract

TFIIA interacts with TFIID via association with TATA binding protein (TBP) and TBP-associated factor 11 (TAF11). We previously identified a mutation in the small subunit of TFIIA (toa2-I27K) that is defective for interaction with TAF11. To further explore the functional link between TFIIA and TAF11, the toa2-I27K allele was utilized in a genetic screen to isolate compensatory mutants in TAF11. Analysis of these compensatory mutants revealed that the interaction between TAF11 and TFIIA involves two distinct regions of TAF11: the highly conserved histone fold domain and the N-terminal region. Cells expressing a TAF11 allele defective for interaction with TFIIA exhibit conditional growth phenotypes and defects in transcription. Moreover, TAF11 imparts changes to both TFIIA-DNA and TBP-DNA contacts in the context of promoter DNA. These alterations appear to enhance the formation and stabilization of the TFIIA-TBP-DNA complex. Taken together, these studies provide essential information regarding the molecular organization of the TAF11-TFIIA interaction and define a mechanistic role for this association in the regulation of gene expression in vivo.

摘要

TFIIA通过与TATA结合蛋白(TBP)和TBP相关因子11(TAF11)相互作用而与TFIID结合。我们之前在TFIIA的小亚基中鉴定出一个突变(toa2-I27K),该突变在与TAF11相互作用方面存在缺陷。为了进一步探索TFIIA与TAF11之间的功能联系,toa2-I27K等位基因被用于遗传筛选,以分离TAF11中的补偿性突变体。对这些补偿性突变体的分析表明,TAF11与TFIIA之间的相互作用涉及TAF11的两个不同区域:高度保守的组蛋白折叠结构域和N端区域。表达与TFIIA相互作用存在缺陷的TAF11等位基因的细胞表现出条件性生长表型和转录缺陷。此外,在启动子DNA的背景下,TAF11会改变TFIIA-DNA和TBP-DNA的接触。这些改变似乎增强了TFIIA-TBP-DNA复合物的形成和稳定性。综上所述,这些研究提供了关于TAF11-TFIIA相互作用分子组织的重要信息,并确定了这种关联在体内基因表达调控中的机制作用。

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