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 是释放新生 RNA 转录本所需的蛋白质 大量革兰氏阴性和革兰氏阴性转录复合物 革兰氏阳性菌。在体外，Rho 结合单链 DNA 和 RNA， 但含胞苷优先激活解旋酶活性 寡核糖核苷酸导致转录终止。自从罗 蛋白质在蛋白质合成的调节中起着核心作用 是细菌代谢的重要且必需的组成部分。 有关原子水平 Rho 核酸相互作用的信息可能 对开发新型抗菌剂具有潜在的帮助。 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研究的主题 经过许多研究人员的深入研究。此外，还有一个大 该蛋白质已进行了多次突变，以努力 了解 Rho 的机制。尽管这项工作导致 我们对 Rho 函数的理解取得了重大进展，这些进展 无法在原子级别解决该机制，因为 没有关于此级别 Rho 结构的信息。 拟议的研究将调查 RNA 结合的结构 Rho 结构域，一个已在大肠杆菌中表达的 15kDa 片段。两个都 NMR 溶液结构和 X 射线晶体结构 该结构域将在各种 RNA 存在和不存在的情况下确定 和DNA寡核苷酸。位点大小和结合常数 DNA 和 RNA 寡核苷酸与 Rho 的该结构域的结合将 评价。结合原子分析此绑定数据 RNA结合域的结构，将为研究提供新的视角 加深对Rho函数的理解。 此外，由于 Rho 包含一本小说 RNA结合基序，这些研究也将极大地增强对RNA结合基序的认识 RNA-蛋白质相互作用的一般情况。