RNA BINDING DOMAIN OF RHO

Rho是一种蛋白质，它是从 革兰氏阴性和阴性的大量转录复合体 革兰氏阳性细菌。在体外，Rho结合单链DNA和RNA， 但含胞苷的解旋酶活性优先激活 寡核苷酸导致转录终止。既然Rho 蛋白质在蛋白质合成的调节中起着核心作用 是细菌新陈代谢的重要组成部分。 有关Rho-核酸在原子水平上相互作用的信息可能 对新型抗菌剂的开发具有潜在的帮助作用。 在过去的十年里，核酸结合特性，酶学， 而这种重要蛋白质的调节特性一直是研究的主题 一些调查人员进行了紧张的研究。此外，一个大型的 已经对这种蛋白质进行了多次突变，以努力 了解RHO的作用机制。尽管这项工作导致了 在我们对Rho函数的理解上取得了重大进展，这些进展 一直无法在原子水平上解决这一机制，因为 在这个层面上，没有关于RHO结构的信息。 拟议的研究将调查RNA结合的结构 Rho的结构域，一个15 kDa的片段，已在大肠杆菌中表达。两者都有 核磁共振溶液结构和X-射线晶体结构 这个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。的站点大小和绑定常数 DNA和RNA寡核苷酸与Rho的这个结构域的结合将是 已评估。分析此绑定数据，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，这些研究也将极大地提高对 一般来说，RNA与蛋白质的相互作用。

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.