Initiation of mRNA decay in Bacillus subtilis

枯草芽孢杆菌中 mRNA 降解的启动

• 批准号: 6470300
• 负责人: DAVID H BECHHOFER
• 金额: $ 32.04万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6470300
• 关键词: Bacillus subtilis; bacterial genetics; chemical cleavage; enzyme activity; exoribonucleases; gene expression; gene mutation; genetic mapping; genetic translation; messenger RNA; microarray technology; microorganism culture; ribonuclease III; Bacillus subtilis; bacterial genetics; chemical cleavage; enzyme activity; exoribonucleases; gene expression; gene mutation; genetic mapping; genetic translation; messenger RNA; microarray technology; microorganism culture; ribonuclease III

DESCRIPTION: (provided by applicant): Our laboratory seeks to understand the control of mRNA decay in Bacillus subtilis. For the Gram-positive bacteria, much needs to be learned about the features of an mRNA that determine its half-life, the ribonucleolytic reactions involved in mRNA decay, the genes that encode ribonuc the regulation of their expression. Since the B. subtilis genome does not have sequence homologues for several of the major ribonuclease genes of Escherichia coli, it is altogether uncertain whether models for mRNA decay based on the well-studied E. coli system will pertain to B. subtilis. Small RNA molecules will be used to probe three facets of mRNA turnover in B. subtilis: 1) the entry site for 3' exonucleolytic degradation; 2) endonucleolytic cleavage that initiates decay; and 3) role of the 5' end in determining mRNA half-life. These small RNA molecules are designed such that analysis of their decay will begin to clarify how mRNA turnover is achieved. To identify additional ribonuclease genes (three have been cloned thus far), biochemical experiments are proposed to isolate several proteins predicted to be involved in mRNA decay, i.e., at least one additional 3'-to-5' exoribonuclease, a putative endoribonuclease, and poly(A) polymerase. Once the identities of these proteins are known, the genes encoding them will be disrupted in order to study the effects on mRNA decay. To develop our understanding of ribonuclease function in the Gram-positive bacteria, we will study the function of Bs-RNase III, a narrow-specificity endoribonuc lease that has been shown to be essential in B. subtilis. Genetic means will be employed in an effort to understand the role of Bs-RNase Ill that is critical for viability. The basis for control of Bs-R.Nase III activity in the cell will be investigated, providing the first look at ribonuclease gene regulation in B. subtilis.

描述：（由申请人提供）：我们的实验室试图了解 控制枯草芽孢杆菌中mRNA衰变。对于革兰氏阳性细菌， 关于mRNA的特征，需要了解很多确定其的特征 半衰期，涉及mRNA衰变的核糖核解反应，该基因 编码核糖表的调节。由于枯草芽孢杆菌基因组 没有几个主要核糖核酸酶基因的序列同源物 大肠杆菌的大肠杆菌，完全不确定mRNA衰变的模型是否 基于研究良好的大肠杆菌系统，将与枯草芽孢杆菌有关。 小的RNA分子将用于探测B中mRNA周转的三个方面。 枯草液：1）3'外核解降解的入口位点； 2） 发起腐烂的内核水解裂解； 3）5'结尾的角色 确定mRNA半衰期。这些小的RNA分子的设计使得 对其衰变的分析将开始阐明如何实现mRNA更新。 为了鉴定其他核糖核酸酶基因（到目前为止已经克隆了三个基因）， 提出了生化实验，以分离预测的几种蛋白质 参与mRNA衰变，即至少另外一个3'-to-5' 驱虫核酸酶，推定的内核酸酶和聚（A）聚合酶。一旦 这些蛋白质的身份是已知的，编码它们的基因将是 为了研究对mRNA衰变的影响而被破坏。 为了发展我们对革兰氏阳性核糖核酸酶功能的理解 细菌，我们将研究BS-RNase III的功能，一个狭窄的特异性 枯草芽孢杆菌中已证明至关重要的内脏租赁。遗传 手段将用于了解BS-RNase病的作用 对于生存能力至关重要。控制BS-R.Nase III活性的基础 该细胞将进行研究，提供核糖核酸酶基因的第一次观察 枯草芽孢杆菌的调节。