Effectors and regulators of end-dependent mRNA degradation in bacteria

细菌末端依赖性 mRNA 降解的效应器和调节器

• 批准号: 8530039
• 负责人: Monica Ping-Ting Hui
• 金额: $ 5.22万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8530039
• 关键词: Address; Affect; Animal Model; Award; Bacillus subtilis; Bacteria; Bacterial Genes; Bacterial Proteins; Biochemical; Biological Assay; Cell physiology; Cells; Degradation Pathway; Enzymes; Escherichia coli; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genetic; Genetic Screening; Glean; Goals; Gram-Positive Bacteria; Growth; Human; Invaded; Investigation; Knowledge; Lead; Life; Measures; Mediator of activation protein; Messenger RNA; Metabolism; Molecular; Nature; Orthologous Gene; Pathogenesis; Pathway interactions; Phase; Phosphorylation; Play; Process; Proteins; RNA; RNA Decay; RNA Degradation; Regulation; Reporter; Ribonucleases; Role; Source; System; Transcript; base; genetic regulatory protein; inorganic phosphate; insight; mRNA Decay; mRNA Transcript Degradation; novel; pathogen; research study; screening; stem; tool

DESCRIPTION (provided by applicant): All bacteria rely on mRNA degradation as a means to regulate gene expression and to adapt to the ever changing milieu in which they live. For a number of transcripts, rapid degradation is governed by the phosphorylation state of the 5' end, which can potentiate RNA decay by nearly an order of magnitude if monophosphorylated. RppH was recently identified as the enzyme responsible for triggering 5' end-dependent degradation by removing phosphates from the 5' end of primary transcripts to produce short-lived 5' monophosphorylated decay intermediates; however, our knowledge of the role of RppH in the larger context of cellular RNA metabolism remains quite rudimentary. The primary purpose of this proposal is to further our understanding of 5' end-dependent RNA decay in bacteria, using Escherichia coli and Bacillus subtilis as model organisms. To achieve this goal, a reporter system will be created to detect changes in 5' end- dependent mRNA degradation, and forward genetics will be used to: 1. identify additional sources of pyrophosphohydrolase activity; and 2. discover positive and negative regulators of RppH. Promising candidates will be further characterized by biochemical and molecular biological assays to determine the scope and mechanism of their influence on 5' end-dependent RNA decay. The results of these studies will enhance our understanding of how bacteria use RNA degradation to control their gene expression profile and provide insights into a regulatory process known to influence bacterial pathogenesis.

描述（由申请人提供）：所有细菌都依赖mRNA降解作为调节基因表达并适应其居住环境不断变化的手段。对于许多转录本，快速降解受5'端的磷酸化状态的控制，如果单磷酸化，可以使RNA衰减几乎增强。最近将RPPH鉴定为负责触发5'末端依赖性降解的酶，通过从主要转录本的5'末端去除磷酸盐，产生短寿命的5'单磷酸化衰减中间体；但是，我们对RPPH在细胞RNA代谢更大背景下的作用的了解仍然是基本的。该提案的主要目的是进一步了解我们对细菌中5'末端依赖性RNA衰变的理解，该RNA使用大肠杆菌和枯草芽孢杆菌作为模型生物。为了实现这一目标，将创建一个记者系统来检测5'最终依赖的mRNA降解中的变化，而正向遗传学将用于：1。识别焦磷酸酶活性的其他来源；和2。发现RPPH的阳性和负调节剂。有希望的候选人将以生化和分子生物学测定的进一步特征，以确定其对5'端依赖性RNA衰变的影响的范围和机制。这些研究的结果将增强我们对细菌如何利用RNA降解来控制其基因表达谱的理解，并提供有关已知影响细菌发病机理的调节过程的见解。