Novel mechanisms of posttranscriptional regulation by CsrA

CsrA转录后调控的新机制

• 批准号: 10326839
• 负责人: Lydia Maria Contreras
• 金额: $ 32万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10326839
• 关键词: 5' Untranslated Regions; Antibiotic Resistance; Bacteria; Bacterial Infections; Binding; Binding Sites; Biological; Biological Assay; Carbon; Cells; Collaborations; Complex; EMSA; Escherichia coli; Fibrinogen; Genes; Genetic Transcription; Goals; Homologous Gene; In Vitro; Knowledge; Lead; Literature; Maps; Messenger RNA; Methods; Modeling; Molecular; Outcome; Pathway interactions; Pattern; Post-Transcriptional Regulation; Proteins; Pseudomonas aeruginosa; RNA; RNA Binding; RNA-Binding Proteins; Regulation; Reporting; Repression; Resistance to infection; Scheme; Stress; System; Testing; Text; Virulence Factors; Work; base; flexibility; genetic regulatory protein; in vivo; innovation; novel; pathogen; prevent; therapeutic target; time use; tool

Abstract (30 lines of text) The mechanisms by which global regulatory proteins operate on their RNA targets are complex and are still largely not understood; understanding these mechanisms is crucial for preventing or alleviating bacterial infections. As one of the major established global post-transcriptional regulator in bacteria, the CsrA RNA- binding protein hides a wealth of strategies for efficient stress-induced posttranscriptional regulation that are not well understood. The overarching goal of the proposed work is to characterize in vivo the molecular features that underlie novel CsrA-target interactions that we have recently uncovered as well as to decipher the biological relevance of these interactions. The CsrA protein is part of the Csr (carbon storage regulator) system, a global regulatory network known to impact virulence factors, secretion systems, and other important genes for rapid host adaptation in a number of pathogens. Based on our recent discoveries, we propose the new hypotheses that: (i) CsrA has diverse types of binding sites on its targets, (ii) CsrA can have multiple binding site combinations within a single target that can lead to different regulatory outcomes, (iii) CsrA interacts with and contributes to the regulation of a wide range of cellular sRNAs, and that (iv) noncanonical features of CsrA regulation are conserved beyond E. coli. Our overall objective is to test these hypotheses in the context of the E. coli CsrA targetome and of the homologue RsmA targetome in P. aeruginosa. Based on our overall goal, we propose three Specific Aims: (1) to characterize in vivo the regulatory binding sites of the CsrA targetome, (2) to characterize in vivo CsrA-sRNA interactions and their functional relevance, and (3) to characterize RsmA-target binding and regulation in P. aeruginosa. We expect that these studies will expand our current paradigm of post-transcriptional regulation approaches that global regulators exert in bacteria. We also expect that this fundamental work will continue to provide framework for studies of CsrA regulation and of other global RNA-binding regulatory proteins to inform their potential use as therapeutic targets. A highly innovative aspect of this work is the use of new intracellular probing tools developed in our lab.

摘要（30行文本） 全局调节蛋白对其RNA靶标的作用机制是复杂的，并且仍然是未知的。 很大程度上还不清楚;了解这些机制对于预防或减轻细菌感染至关重要。 感染.作为细菌中主要的转录后调节因子之一，CsrA RNA- 结合蛋白隐藏了大量的策略，有效的压力诱导转录后调节， 没有很好地理解。拟议工作的首要目标是在体内表征分子 这些特征是我们最近发现的新的CsrA-靶相互作用的基础， 这些相互作用的生物相关性。CsrA蛋白是Csr（碳储存调节剂）的一部分。 系统，一个已知影响毒力因子、分泌系统和其他 在许多病原体中快速适应宿主的重要基因。根据我们最近的发现，我们 提出了新的假设：（i）CsrA在其靶点上具有不同类型的结合位点，（ii）CsrA可以具有 单个靶标内的多个结合位点组合，其可导致不同的调控结果，（iii） CsrA与广泛的细胞sRNA相互作用并促进其调节，并且（iv） CsrA调控的非典型特征在E.杆菌我们的总体目标是测试这些 在E. coliCsrA targetome和P. 铜绿。基于我们的总体目标，我们提出了三个具体目标：（1）在体内表征 CsrA靶向组的调节结合位点，（2）表征体内CsrA-sRNA相互作用及其 功能相关性，和（3）表征铜绿假单胞菌中的RsmA-靶标结合和调节。我们 我希望这些研究将扩展我们目前的转录后调控方法的范式， global全球regulators调节effects施加in bacteria细菌.我们还期望这项基础性工作将继续提供 CsrA调控和其他全球RNA结合调控蛋白的研究框架，以告知他们的 作为治疗靶点的潜在用途。这项工作的一个高度创新的方面是使用新的细胞内 我们实验室开发的探测工具