The transcription elongation factor TFIIS is a component of RNA polymerase II preinitiation complexes

被引:87
作者
Kim, Bong
Nesvizhskii, Alexey I.
Rani, P. Geetha
Hahn, Steven
Aebersold, Ruedi
Ranish, Jeffrey A.
机构
[1] Inst Syst Biol, Seattle, WA 98103 USA
[2] Univ Michigan, Dept Pathol, Ann Arbor, MI 48109 USA
[3] Fred Hutchinson Canc Res Ctr, Div Basic Sci, Seattle, WA 98109 USA
[4] ETH, Inst Mol Syst Biol, CH-8092 Zurich, Switzerland
[5] Univ Zurich, Fac Sci, CH-8006 Zurich, Switzerland
关键词
isotope-coded affinity tagging; quantitative mass spectrometry; Saccharomyces cerevisiae; stable isotopes;
D O I
10.1073/pnas.0704573104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
In this article, we provide direct evidence that the evolutionarily conserved transcription elongation factor THIS functions during preinitiation complex assembly. First, we identified THIS in a mass spectrometric screen of RNA polymerase II (PoI II) preinitiation complexes (PICs). Second, we show that the association of THIS with a promoter depends on functional PIC components including Mediator and the SAGA complex. Third, we demonstrate that THIS is required for efficient formation of active PICs. Using truncation mutants of THIS, we find that the PoI II-binding domain is the minimal domain necessary to stimulate PIC assembly. However, efficient formation of active PICs requires both the PoI II-binding domain and the poorly understood N-terminal domain. Importantly, Domain III, which is required for the elongation function of THIS, is dispensable during PIC assembly. The results demonstrate that TFIIS is a PIC component that is required for efficient formation and/or stability of the complex.
引用
收藏
页码:16068 / 16073
页数:6
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