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RNA BINDING DOMAIN OF RHO

RHO 的 RNA 结合域

基本信息

批准号：
2750142
负责人：
GORDON S. RULE
金额：
$ 16.17万
依托单位：
CARNEGIE-MELLON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-08-01 至 1999-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2750142
关键词：
RNA binding protein X ray crystallography bacterial proteins chemical binding circular dichroism intermolecular interaction molecular dynamics molecular site mutant nuclear magnetic resonance spectroscopy oligonucleotides protein structure site directed mutagenesis transcription factor

项目摘要

Rho is a protein required for the release of nascent RNA transcripts from a large number of transcription complexes in both gram-negative and gram-positive bacteria. In vitro, Rho binds single stranded DNA and RNA, but preferential activation of helicase activity by cytidine-containing oligoribonucleotides leads to termination of transcription. Since Rho protein plays a central role in the regulation of protein synthesis it is a significant and essential component of bacterial metabolism. Information about Rho-nucleic acid interactions at the atomic level may be of potential aid in the development of novel anti-bacterial agents. Within the past decade the nucleic acid binding properties, enzymology, and regulatory properties of this important protein have been the subject of intense studies by a number of investigators. Additionally, a large number of mutations of this protein have been made in an effort to understand the mechanism of Rho. Although this work has lead to significant advances in our understanding of Rho function, these advances have not been able to address the mechanism at the atomic level because there is no information on the structure of Rho at this level. The proposed research will investigate the structure of the RNA binding domain of Rho, a 15kDa fragment which has been expressed in E. coli. Both the NMR solution structure and the x-ray crystallographic structure of this domain will be determined in the presence and absence of various RNA and DNA oligonucleotides. The site size and binding constant for the binding of DNA and RNA oligonucleotides to this domain of Rho will be evaluated. Analysis of this binding data, in conjunction with the atomic structure of the RNA binding domain, will provide a new perspective in the understanding of Rho function. Further, since Rho contains a novel RNA-binding motif, these studies will also greatly enhance knowledge of RNA-protein interactions in general.

Rho是一种蛋白质，用于从细胞中释放新生RNA转录物。大量的转录复合物在革兰氏阴性和革兰氏阳性菌在体外，Rho结合单链DNA和RNA，但通过含胞苷的寡核苷酸优先激活解旋酶活性寡核糖核苷酸导致转录终止。自从Rho 蛋白质在蛋白质合成的调节中起着核心作用，是细菌代谢的重要组成部分。在原子水平上关于Rho-核酸相互作用的信息可以可能有助于开发新型抗菌剂。在过去的十年中，核酸结合特性，酶学，这种重要蛋白质的调节特性一直是许多研究者的密集研究。此外，一个大这种蛋白质的许多突变已经被进行了努力，了解Rho的机制。虽然这项工作已经导致我们对Rho功能的理解有了重大进展，这些进展还没有能够在原子水平上解决这个机制，没有关于Rho在这一级的结构的信息。拟议的研究将调查RNA结合的结构， Rho结构域的一个15 kDa的片段，已在E.杆菌两 NMR溶液结构和X射线晶体结构该结构域将在存在和不存在各种RNA的情况下确定，和DNA寡核苷酸。的位点大小和结合常数 DNA和RNA寡核苷酸与Rho的该结构域的结合将是评估。分析这个绑定数据，结合原子 RNA结合域的结构，将提供一个新的视角，对Rho函数的理解。此外，由于Rho包含一部小说， RNA结合基序，这些研究也将大大提高知识， RNA与蛋白质的相互作用。