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: 8312988
• 负责人: Monica Ping-Ting Hui
• 金额: $ 4.92万
• 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/8312988
• 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; Screening procedure; Source; System; Transcript; base; genetic regulatory protein; inorganic phosphate; insight; mRNA Decay; mRNA Transcript Degradation; novel; pathogen; research study; 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. PUBLIC HEALTH RELEVANCE: RppH is a bacterial protein previously implicated as being important for pathogens to invade human cells. Furthermore, all bacteria-non-pathogenic and pathogenic alike-rely on mRNA degradation to regulate gene expression and to adapt to their surroundings. These studies will enhance and deepen our understanding of a fundamental cellular process, thereby providing insight into a molecular mechanism 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和适应不断变化的生存环境的手段。对于许多转录物，快速降解由5'端的磷酸化状态控制，如果单磷酸化，其可以增强RNA衰变近一个数量级。RppH最近被确定为负责触发5'末端依赖性降解的酶，通过从初级转录物的5'末端去除磷酸盐以产生短寿命的5'单磷酸化衰变中间体;然而，我们对RppH在细胞RNA代谢的更大背景下的作用的了解仍然相当基本。本研究的主要目的是以大肠杆菌和枯草芽孢杆菌为模式生物，进一步了解细菌中5'末端依赖的RNA衰变。为了实现这一目标，将创建报告系统来检测5'末端依赖性mRNA降解的变化，并且将使用正向遗传学来：1.鉴定焦磷酸水解酶活性的其它来源;和2.发现RppH的正负调节因子。有希望的候选物将通过生物化学和分子生物学测定进一步表征，以确定它们对5'末端依赖性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，并提供对已知影响细菌发病机制的调控过程的见解。 公共卫生相关性：RppH是一种细菌蛋白，以前被认为是病原体入侵人体细胞的重要因素。此外，所有细菌-非致病性和致病性-都依赖于mRNA降解来调节基因表达和适应环境。这些研究将增强和加深我们对基本细胞过程的理解，从而深入了解已知影响细菌发病机制的分子机制。