RNA STRUCTURE IN AGILE LOCI OF RIBOSOME AND RETROVIRUSES

核糖体和逆转录病毒敏捷位点的 RNA 结构

• 批准号: 6499498
• 负责人: THOMAS L JAMES
• 金额: $ 3.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-09 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6499498
• 关键词: DNA; Retroviridae; antisense nucleic acid; bacterial RNA; chemical stability; conformation; cooperative study; dimer; double stranded RNA; endoribonucleases; gel mobility shift assay; gene expression; genetic promoter element; genetic regulation; molecular dynamics; nuclear magnetic resonance spectroscopy; nucleic acid structure; oligonucleotides; protein biosynthesis; ribosomal RNA; ribosomes; solutions; statistics /biometry; virus RNA

A collaborative project is proposed between UCSF and the Engelhardt Institute of Molecular Biology, Russian Academy of Science, Moscow, in the area of RNA conformations. The parent grant for the present proposal is GM39247 (7/1/98-6/30/02). Alternative conformations of RNA will be studied for functionally important sequences. Specifically, two cases will be investigated where conformation switching of RNA may play an important functional role: (1) structures that are formed during the dimerization of genomic RNA of avian retroviruses. (2) a hypothetical transient RNA psuedoknot at the ribosomal peptidyl transferase cancer, proposed recently by the Moscow group based on phylogenetic analysis of ribosomal RNA sequences. A set of RNA oligonucleotides (40 to 50 nucleotides in length) will be studied in solution, including sequences from a number of avian retroviruses, human and bacterial rRNA sequences, a number of mutated sequences. In this project, the Moscow group will characterize the RNA oligonucleotides by physicochemical methods and map the single- and double-stranded regions using chemicals and enzymatic probing. Based on these data, three-dimensional models will be calculated for the structures involved, and suitable sequences will be studied by UCSF groups using high resolution NMR. A reversible formation of a pseudoknot structure at certain stages of the ribosome operation may be responsible for the large-scale dynamics during protein synthesis. Understanding the detailed mechanisms of this process as well as that of the retroviral dimerization and knowledge of the structures involved may open new perspectives in designing new bacterial and antiviral therapeutic agents.

在RNA构象领域，俄罗斯科学学院，俄罗斯科学学院的恩格哈特分子生物学研究所之间提出了一个合作项目。本提案的父母赠款为GM39247（7/1/98-6/30/02）。将研究RNA的替代构象以进行功能重要的序列。具体而言，将研究两种情况，其中RNA的构象转换可能起着重要的功能作用：（1）在鸟类逆转录病毒基因组RNA二聚化过程中形成的结构。 （2）核糖体肽基转移酶癌症的假设瞬态RNA psuedoknot，最近由莫斯科组基于核糖体RNA序列的系统发育分析提出。将在溶液中研究一组RNA寡核苷酸（长度为40至50个核苷酸），包括来自许多禽逆转录病毒，人类和细菌RRNA序列的序列，许多突变的序列。在该项目中，莫斯科组将通过理化方法表征RNA寡核苷酸，并使用化学物质和酶探测绘制单链和双链区域。基于这些数据，将针对所涉及的结构计算三维模型，并使用高分辨率NMR对UCSF组研究合适的序列。在核糖体操作的某些阶段，伪诺结构的可逆形成可能是蛋白质合成过程中大规模动力学的原因。了解此过程的详细机制以及逆转录病毒二聚化和所涉及结构知识的详细机制可能会在设计新的细菌和抗病毒治疗剂时开放新的观点。