YloC, a new ribonuclease of Bacillus subtilis

YloC,枯草芽孢杆菌的新型核糖核酸酶

基本信息

项目摘要

PROJECT SUMMARY: Our laboratory has, for many years, studied the essential process of mRNA decay in the model Gram-positive bacterium, Bacillus subtilis. We have identified several ribonuclease (RNase) enzymes of B. subtilis and have elucidated the role they play in mRNA turnover. The viability of a B. subtilis strain lacking all of the known 3’-to-5’ exoribonucleases prompted us to pursue identification of additional RNase activities. Using classic protein biochemistry, we recently identified a novel RNase, named YloC. YloC is an endoribonuclease with a hexameric structure, an unusual characteristic that is shared with only one other RNase: the Nsp15 protein of the SARS-CoV family. Initial experiments suggest that, although YloC has ribonuclease activity in vitro, it may function as an adapter for RNA interactions in vivo. Although proteins with significant homology to YloC are widespread in bacterial species, there is no published information on the structure of any member of this protein family. The current proposal seeks to elucidate the structure and function of YloC, as follows: • Mutagenize highly conserved residues of YloC to determine the effect on several properties – including ribonuclease activity, RNA binding, and structure – and to clarify functional domains of the protein. • Identify high-affinity RNA ligands of YloC via SELEX procedures with random-sequence oligonucleotides and with genomic RNA sequences. • Characterize how the strong interaction of YloC with E. coli polynucleotide phosphorylase (PNPase) acts in small RNA (sRNA) regulation in E. coli and possibly in B. subtilis. • Determine the three-dimensional structure of the highly homologous E. coli YicC protein bound to an RNA substrate, as well as the structure of YloC and/or its homologs from thermophilic bacterial species. This work will build on an initial determination of the structure of YicC. RELEVANCE: Ribonucleases play essential roles in RNA turnover and processing. A thorough understanding of the proteins that bind to and act enzymatically on RNA molecules will enable design of antimicrobial agents that disrupt such proteins and thereby interfere with bacterial cell growth.
项目概述:我们的实验室多年来一直在研究mRNA衰变的基本过程, 革兰氏阳性菌枯草芽孢杆菌。我们已经鉴定了几种核糖核酸酶 的B。枯草芽孢杆菌,并阐明了它们在mRNA周转中的作用。B的生存能力。缺乏枯草杆菌菌株 所有已知的3 ′-至5 ′核糖核酸外切酶促使我们继续鉴定另外的RNA酶活性。 使用经典的蛋白质生物化学,我们最近发现了一种新的RNase,命名为YloC。YLoC是一个 具有六聚体结构的核糖核酸内切酶,这是一种仅与另一种RNA酶共享的不寻常特征: SARS-CoV家族的Nsp 15蛋白。最初的实验表明,尽管YloC具有核糖核酸酶, 在体外的活性,它可以作为一个适配器的RNA相互作用在体内。虽然蛋白质具有显著的 尽管与YloC的同源性在细菌物种中广泛存在,但是没有关于任何YloC的结构的公开信息, 这个蛋白质家族的成员。目前的建议旨在阐明YLoC的结构和功能, 如下所示: ·诱变YloC的高度保守残基以确定对几种性质的影响,包括 核糖核酸酶活性,RNA结合和结构-并阐明蛋白质的功能结构域。 ·通过具有随机序列的SELEX程序鉴定YloC的高亲和力RNA配体 寡核苷酸和基因组RNA序列。 ·表征YloC与E.大肠杆菌多核苷酸磷酸化酶 在E. coli中,可能存在于B.枯草芽孢杆菌 ·确定高度同源的E.大肠杆菌YicC蛋白结合到一个 RNA底物,以及来自嗜热细菌物种的YloC和/或其同源物的结构。 这项工作将建立在YicC结构的初步确定。 相关性:核糖核酸酶在RNA周转和加工中发挥重要作用。的透彻理解 与RNA分子结合并对RNA分子起酶促作用的蛋白质将使抗菌剂的设计成为可能 破坏这些蛋白质,从而干扰细菌细胞生长。

项目成果

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DAVID H BECHHOFER其他文献

DAVID H BECHHOFER的其他文献

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{{ truncateString('DAVID H BECHHOFER', 18)}}的其他基金

Global analysis of mRNA decay in Bacillus subtilis
枯草芽孢杆菌 mRNA 衰减的整体分析
  • 批准号:
    8655171
  • 财政年份:
    2012
  • 资助金额:
    $ 35.49万
  • 项目类别:
Global analysis of mRNA decay in Bacillus subtilis
枯草芽孢杆菌 mRNA 衰减的整体分析
  • 批准号:
    8515472
  • 财政年份:
    2012
  • 资助金额:
    $ 35.49万
  • 项目类别:
Global analysis of mRNA decay in Bacillus subtilis
枯草芽孢杆菌 mRNA 衰减的整体分析
  • 批准号:
    8371861
  • 财政年份:
    2012
  • 资助金额:
    $ 35.49万
  • 项目类别:
Initiation of mRNA decay in Bacillus subtilis
枯草芽孢杆菌中 mRNA 降解的启动
  • 批准号:
    7921228
  • 财政年份:
    2009
  • 资助金额:
    $ 35.49万
  • 项目类别:
Initiation of mRNA decay in Bacillus subtilis
枯草芽孢杆菌中 mRNA 降解的启动
  • 批准号:
    6470300
  • 财政年份:
    1993
  • 资助金额:
    $ 35.49万
  • 项目类别:
Initiation of mRNA decay in Bacillus subtilis
枯草芽孢杆菌中 mRNA 降解的启动
  • 批准号:
    7092749
  • 财政年份:
    1993
  • 资助金额:
    $ 35.49万
  • 项目类别:
Initiation of mRNA decay in Bacillus subtilis
枯草芽孢杆菌中 mRNA 降解的启动
  • 批准号:
    7629564
  • 财政年份:
    1993
  • 资助金额:
    $ 35.49万
  • 项目类别:
INITIATION OF MRNA DECAY IN BACILLUS SUBTILIS
枯草芽孢杆菌中 mRNA 衰变的启动
  • 批准号:
    2186314
  • 财政年份:
    1993
  • 资助金额:
    $ 35.49万
  • 项目类别:
INITIATION OF MRNA DECAY IN BACILLUS SUBTILIS
枯草芽孢杆菌中 mRNA 衰变的启动
  • 批准号:
    2022688
  • 财政年份:
    1993
  • 资助金额:
    $ 35.49万
  • 项目类别:
Initiation of mRNA decay in Bacillus subtilis
枯草芽孢杆菌中 mRNA 降解的启动
  • 批准号:
    8073263
  • 财政年份:
    1993
  • 资助金额:
    $ 35.49万
  • 项目类别:

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冠状病毒外核糖核酸酶复合物多方面作用的机制见解
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