Molecular Mechanisms of mRNA Turnover

mRNA 周转的分子机制

• 批准号: 8917102
• 负责人: Janet Schottel
• 金额: $ 25.5万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-15 至 1993-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8917102&HistoricalAwards=false
• 关键词: Molecular; Mechanisms; mRNA; Turnover

The decay rate of a particular mRNA can greatly influence the amount of protein produced from that message and can thereby play an important role in the overall regulation of gene expression. The goal of this project is to elucidate the mechanisms by which cells degrade messenger RNA molecules and are able to regulate this decay process. The proposed experiments will determine the mechanism by which the E. coli chloramphenicol acetyltransferase (cat) message is degraded and how the stability of this message is regulated in response to culture growth conditions. Three cleavage sites have been detected within the cat message that may be involved in initiating the decay process. These cleavage sites will be mapped, and the importance of these cleavages to the decay process will be determined by deletion and site-directed mutagenesis and by gene fusion analyses. An in vitro message degradation assay will be used to study the decay process and to eventually purify the ribonucleases involved in these degradative events. Changes in the stability of the cat message in response to non-steady state growth conditions and medium composition will also be measured and compared to other messages such as ompA, bla, lpp, lac, and trxA. The results of these experiments will define the important components of the cat message decay pathway, which should be broadly applicable to other messages in E. coli, and will be lend insight into how message stability is regulated in response to culture growth conditions.

特定mRNA的衰变率可以极大地影响细胞的增殖。 从信息中产生的蛋白质的数量， 在基因表达的整体调控中起重要作用。 本项目的目标是阐明 细胞降解信使RNA分子， 这个腐烂的过程。 拟议的实验将确定 E.大肠杆菌氯霉素乙酰转移酶 (cat)消息被降级，以及此消息的稳定性 是受培养物生长条件调节的。 三 在CAT信息中检测到切割位点， 参与了衰变过程的启动 这些切割 这些裂缝的重要性， 衰变过程将由删除决定， 定点诱变和基因融合分析。 的in 体外信息降解试验将用于研究衰变 处理并最终纯化参与的核糖核酸酶， 这些退化的事件。 猫的稳定性变化 响应于非稳态生长条件的消息，以及 还将测量培养基成分并与其他培养基成分进行比较。 消息，例如ompA、bla、lpp、lac和trxA。 的结果 这些实验将确定猫的重要组成部分 消息衰减路径，这应该广泛适用于 E.大肠杆菌，并将有助于了解如何 消息稳定性是根据文化的发展而调节的 条件