Transcription activation by a sliding clamp.

Transcription activation by a sliding clamp.
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DOI:
10.1038/s41467-021-21392-0
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发表时间:
2021-02-18
影响因子:
16.6
通讯作者:
Feng Y
Feng Y
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Shi J;Wen A;Jin S;Gao B;Huang Y;Feng Y

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噬菌体T4晚期基因的转录激活是通过包含RNA聚合酶(RNAP)、启动子特异性因子gp 55、共激活因子gp 33和细胞DNA复制的通用组分滑动夹gp 45的转录激活复合物来完成的。虽然遗传和生物化学研究已经阐明了T4晚期基因转录的许多方面,但该过程中的转录机制没有精确的结构。在这里,我们报告的冷冻电镜结构的gp 55依赖RNAP启动子开放复合物和完整的gp 45依赖的转录激活复合物。这些结构揭示了gp 55与介导T4晚期启动子识别的启动子DNA之间的相互作用。除了σR2同源结构域之外,gp 55还具有螺旋-环-螺旋基序,其陪伴转录泡的模板链单链DNA。Gp 33接触RNAP和上游双链DNA。Gp 45包围DNA并将RNAP与其连接,支持gp 45将启动子搜索从三维扩散模式切换到一维扫描模式的想法。T4噬菌体中晚期基因的转录激活需要启动子特异性因子gp 55、共激活因子gp 33和滑动夹gp 45。在这里,作者通过确定gp 55依赖性RNA聚合酶(RNAP)启动子开放复合物和完整的gp 45依赖性转录激活复合物的冷冻EM结构,提供了对gp 45依赖性转录激活的结构见解。
Transcription activation of bacteriophage T4 late genes is accomplished by a transcription activation complex containing RNA polymerase (RNAP), the promoter specificity factor gp55, the coactivator gp33, and a universal component of cellular DNA replication, the sliding clamp gp45. Although genetic and biochemical studies have elucidated many aspects of T4 late gene transcription, no precise structure of the transcription machinery in the process is available. Here, we report the cryo-EM structures of a gp55-dependent RNAP-promoter open complex and an intact gp45-dependent transcription activation complex. The structures reveal the interactions between gp55 and the promoter DNA that mediate the recognition of T4 late promoters. In addition to the σR2 homology domain, gp55 has a helix-loop-helix motif that chaperons the template-strand single-stranded DNA of the transcription bubble. Gp33 contacts both RNAP and the upstream double-stranded DNA. Gp45 encircles the DNA and tethers RNAP to it, supporting the idea that gp45 switches the promoter search from three-dimensional diffusion mode to one-dimensional scanning mode. Transcription activation of late genes in T4 bacteriophage requires the promoter specificity factor gp55, the coactivator gp33 and the sliding clamp gp45. Here, the authors provide structural insights into gp45- dependent transcription activation by determining the cryo-EM structures of a gp55-dependent RNA polymerase (RNAP)-promoter open complex and of an intact gp45-dependent transcription activation complex.
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