RNA BINDING DOMAIN OF RHO

RHO 的 RNA 结合域

• 批准号: 2459733
• 负责人: GORDON S. RULE
• 金额: $ 16.82万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 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/2459733
• 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

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是从中释放新生RNA转录本所需的蛋白 革兰氏阴性和 革兰氏阳性细菌。在体外，rho结合了单链DNA和RNA， 但是通过含胞苷的旋转酶活性优先激活 寡核苷酸导致转录终止。自Rho以来 蛋白质在调节蛋白质合成中起着核心作用 是细菌代谢的重要组成部分。 关于原子水平的Rho-核酸相互作用的信息可能 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， 该重要蛋白质的调节特性一直是受试者 许多研究者的深入研究。另外，一个大 为了努力，该蛋白的突变数量是为了 了解Rho的机制。尽管这项工作已导致 这些进步在我们对Rho功能的理解方面取得了重大进展 无法在原子层面上解决该机制，因为 在此级别上没有关于Rho结构的信息。 拟议的研究将研究RNA结合的结构 Rho的域，这是一种15kDa碎片，已在大肠杆菌中表达。两个都 NMR溶液结构和X射线晶体学结构 该域将在存在和不存在各种RNA的情况下确定 和DNA寡核苷酸。位点大小和结合常数 DNA和RNA寡核苷酸与该RHO域的结合将是 评估。分析该结合数据，并结合原子 RNA结合域的结构将提供一个新的视角 对Rho功能的理解。 此外，由于Rho包含小说 RNA结合图案，这些研究还将大大增强对 RNA蛋白相互作用一般。