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

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）：我们的实验室旨在了解 枯草芽孢杆菌中mRNA衰变的控制。对于革兰氏阳性菌， 需要了解更多关于mRNA的特征，这些特征决定了其 半衰期，参与mRNA衰变的核糖核酸溶解反应， 编码核糖核酸调控其表达。从B开始。枯草杆菌基因组 没有几个主要核糖核酸酶基因的序列同源物 对于大肠杆菌，mRNA衰变的模型是否完全不确定 基于已被广泛研究的E. coli系统将属于B。枯草杆菌。 小RNA分子将用于探测B中mRNA周转的三个方面。 枯草芽孢杆菌：1）3'核酸外切降解的进入位点; 2） 启动衰变的核酸内切裂解;和3）5'末端在 确定mRNA半衰期。这些小RNA分子被设计成 将开始对它们的衰变进行分析，以阐明mRNA周转是如何实现的。 为了鉴定另外的核糖核酸酶基因（迄今为止已经克隆了三个）， 提出了生物化学实验来分离几种预测为 参与mRNA衰变，即，至少一个额外的3“至5” 核糖核酸外切酶、推定的核糖核酸内切酶和聚腺苷酸聚合酶。一旦 这些蛋白质的身份是已知的，编码它们的基因将是 破坏以研究对mRNA衰变的影响。 为了加深我们对核糖核酸酶在革兰氏阳性 细菌，我们将研究Bs-RNase III的功能， 在B中已被证明是必不可少的。枯草杆菌。遗传 将采用各种方法来努力理解Bs-RNase III的作用， 对生存能力至关重要控制Bs-R.Nase III活性的基础 细胞将被调查，提供第一次看核糖核酸酶基因 B中的调节。枯草杆菌。