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首页> 外文学位 >Dissecting the interaction between Escherichia coli 6S RNA and the RNA polymerase sigma70 subunit.
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Dissecting the interaction between Escherichia coli 6S RNA and the RNA polymerase sigma70 subunit.

机译：解剖大肠杆菌6S RNA和RNA聚合酶sigma70亚基之间的相互作用。

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摘要

The Escherichia coli 6S RNA is a small RNA which forms a specific, stable complex with RNA Polymerase (RNAP) containing sigma 70 (Esigma70). sigma70 is the RNAP subunit that conveys specificity to the enzyme by making direct contact with the -35 and -10 hexamers that define sigma70-dependent promoters. sigma 70 contains four conserved regions that each play a role in contacting promoter elements, including region 2 binding to the -10 hexamer and region 4 binding to the -35 hexamer.;6S RNA forms a phylogenetically conserved secondary structure that resembles promoter DNA in the open complex conformation, and this secondary structure is essential for the interaction with Esigma70. By binding Esigma70, 6S RNA inhibits transcription from a subset of sigma 70-dependent promoters by directly competing with promoter DNA for binding to Esigma70. 6S RNA is found within the Esigma 70 active site, suggesting that the 6S RNA-Esigma70 contacts overlap the location on Esigma70 that binds promoter DNA.;My thesis research was to further understand the 6S RNA-Esigma 70 interaction by analyzing sigma70 mutants to elucidate a 6S RNA binding surface onsigma70 (within Esigma 70). I have shown that sigma70 region 4.2, which contacts the -35 hexamer through a helix-turn-helix structural motif, is critical for binding 6S RNA as well. sigma70 subunits with single alanine substitutions identified residues within sigma70 region 4.2 that are important for binding 6S RNA. These studies showed that the 6S RNA binding surface on sigma70 region 4.2 is distinct but overlapping with the DNA binding site. Furthermore, the 6S RNA binding surface is defined by multiple positively-charged residues, all of which contribute significantly to binding. My research has defined an RNA binding interaction with sigma70 region 4.2 that is divergent from DNA binding, which is likely to be one of many contacts between 6S RNA and Esigma70.
机译：大肠杆菌6S RNA是小的RNA，可与包含sigma 70（Esigma70）的RNA聚合酶（RNAP）形成特异性，稳定的复合物。 sigma70是RNAP亚基，通过与定义sigma70依赖启动子的-35和-10六聚体直接接触，向酶传达特异性。 sigma 70包含四个保守区域，每个区域均在启动子元件接触中发挥作用，包括与-10六聚体结合的区域2和与-35六聚体结合的区域4.; 6S RNA形成系统发育上保守的二级结构，类似于启动子DNA。开放的复杂构象，并且这种二级结构对于与Esigma70的相互作用至关重要。通过结合Esigma70，6S RNA通过直接与启动子DNA竞争与Esigma70的结合而抑制了来自sigma 70依赖性启动子的转录。在Esigma 70活性位点中发现6S RNA，这表明6S RNA-Esigma70接触与结合启动子DNA的Esigma70重叠。我的论文研究是通过分析sigma70突变体来进一步了解6S RNA-Esigma 70相互作用。 6S RNA结合表面onsigma70（在Esigma 70内）。我已经证明，通过一个螺旋-转-螺旋结构基序与-35六聚体接触的sigma70区4.2对于结合6S RNA也至关重要。具有单个丙氨酸取代的sigma70亚基鉴定出sigma70区域4.2中的残基对于结合6S RNA很重要。这些研究表明，在sigma70区域4.2上的6S RNA结合表面是不同的，但与DNA结合位点重叠。此外，6S RNA结合表面由多个带正电荷的残基定义，所有这些均显着促进结合。我的研究定义了与sigma70区4.2的RNA结合相互作用，该相互作用不同于DNA结合，这很可能是6S RNA和Esigma70之间的许多接触之一。

著录项

  • 作者

    Klocko, Andrew D.;

  • 作者单位

    The University of Wisconsin - Madison.;

  • 授予单位 The University of Wisconsin - Madison.;
  • 学科 Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 155 p.
  • 总页数 155
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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