Divergent contributions of conserved active site residues to transcription by eukaryotic RNA polymerases I and II.

Divergent contributions of conserved active site residues to transcription by eukaryotic RNA polymerases I and II.
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DOI:
10.1016/j.celrep.2013.07.044
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发表时间:
2013-09-12
期刊:
影响因子:
8.8
通讯作者:
Schneider DA
Schneider DA
中科院分区:
生物学1区
文献类型:
--
作者:
Viktorovskaya OV;Engel KL;French SL;Cui P;Vandeventer PJ;Pavlovic EM;Beyer AL;Kaplan CD;Schneider DA

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多亚基RNA聚合酶(msRNAP)具有高度的序列和结构同源性,特别是在其活性位点内,这通常被认为是导致msRNAP功能保守性的原因。然而,我们发现,突变的触发环(TL)在RNA聚合酶I(Pol I)的最大亚基产生的表型出乎意料的Pol II的研究。例如,Pol II中充分表征的功能获得性突变导致Pol I中的功能丧失[Pol II:rpb 1-E1103 G; Pol I:rpa 190-E1224 G]。嵌合Pol II酶托管Pol I或Pol III TL的研究表明,改变TL动力学的突变的后果是由更大的酶环境决定的,而不仅仅是TL序列。尽管rpa 190-E1224 G突变降低了聚合酶功能,但当与干扰Pol I催化的突变结合时,它增强了聚合酶功能,类似于类似的Pol II突变。这些结果表明,Pol I和Pol II具有不同的限速步骤。
Multisubunit RNA polymerases (msRNAPs) exhibit high sequence and structural homology, especially within their active sites, which is generally thought to result in msRNAP functional conservation. However, we show that mutations in the trigger loop (TL) in the largest subunit of RNA polymerase I (Pol I) yield phenotypes unexpected from studies of Pol II. For example, a well-characterized gain-of-function mutation in Pol II results in loss-of-function in Pol I [Pol II: rpb1- E1103G; Pol I: rpa190-E1224G]. Studies of chimeric Pol II enzymes hosting Pol I or Pol III TLs suggest that consequences of mutations that alter TL dynamics are dictated by the greater enzymatic context and not solely the TL sequence. Although the rpa190-E1224G mutation diminishes polymerase function, when combined with mutations that perturb Pol I catalysis, it enhances polymerase function, similar to the analogous Pol II mutation. These results suggest that Pol I and Pol II have different rate-limiting steps.
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