Structural basis of initial RNA polymerase II transcription.

Structural basis of initial RNA polymerase II transcription.
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DOI:
10.1038/emboj.2011.396
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发表时间:
2011-11-04
期刊:
影响因子:
11.4
通讯作者:
Cramer, Patrick
Cramer, Patrick
中科院分区:
生物学1区
文献类型:
--
作者:
Cheung, Alan C. M.;Sainsbury, Sarah;Cramer, Patrick

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在RNA聚合酶(Pol)II的转录起始过程中,瞬时开放启动子复合体(OC)被转化为含有短RNA的初始转录复合体(ITC),并转化为稳定的延伸复合体(EC)。我们报道了Pol II-DNA复合体的结构,该复合体模拟OC的一部分,以及代表最小ITCs的复合体的结构,该复合体具有2,4,5,6和7个核苷酸(NT)RNA,在插入位点+1中有和没有不可水解的核苷三磷酸(NTP)。部分OC结构表明POL II将熔融的模板链定位在活性中心的对面。模拟ITC的结构表明,两个不变的赖氨酸残基锚定了短RNA的3‘-近端磷酸。短DNA-RNA杂交物采用倾斜构象,将+1模板NT从活性部位排除在外。NTP结合可诱导完整的DNA易位和标准杂交构象。保守的NTP联系人表明了NTP选择的通用机制。闭合触发环中的基本残基Q1078与NTP 2‘-OH基团结合,解释了触发环如何将催化与NTP选择偶联,抑制dNTP结合和DNA合成。
During transcription initiation by RNA polymerase (Pol) II, a transient open promoter complex (OC) is converted to an initially transcribing complex (ITC) containing short RNAs, and to a stable elongation complex (EC). We report structures of a Pol II–DNA complex mimicking part of the OC, and of complexes representing minimal ITCs with 2, 4, 5, 6, and 7 nucleotide (nt) RNAs, with and without a non-hydrolyzable nucleoside triphosphate (NTP) in the insertion site +1. The partial OC structure reveals that Pol II positions the melted template strand opposite the active site. The ITC-mimicking structures show that two invariant lysine residues anchor the 3′-proximal phosphate of short RNAs. Short DNA–RNA hybrids adopt a tilted conformation that excludes the +1 template nt from the active site. NTP binding induces complete DNA translocation and the standard hybrid conformation. Conserved NTP contacts indicate a universal mechanism of NTP selection. The essential residue Q1078 in the closed trigger loop binds the NTP 2′-OH group, explaining how the trigger loop couples catalysis to NTP selection, suppressing dNTP binding and DNA synthesis.
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