Probing the Metabolism and Cellular Roles of 2',3'-Cyclic Nucleotide Monophosphate

摘要 新型细胞内小分子2‘，3’-环核苷酸单磷酸(2‘，3’-cNMPs) 在原核生物和真核生物中都被发现。在植物和哺乳动物中，伤害一直是 发现会导致2‘，3’-cNMPs水平升高。对细菌的初步研究表明，2‘，3’-cNMP 2‘，3’-cNMP水平会影响细菌的表型，如 如生物膜的形成和运动，以及众多基因的表达。这项研究的长期目标是 了解2‘，3’-cNMPs在控制原核生物信号通路中的作用并加以利用 设计小分子来调节细胞表型的知识。这一目标将通过以下方式实现 研究参与2‘，3’-cNMP代谢的细胞成分，鉴定2‘，3’-cNMP感受器， 以及确定2‘，3’-cNMPs对多种细菌下游途径的影响。这个 拟议的工作概述了一项创新的研究计划，以探索一种新的细胞应力传感机制和 将提供有关2‘，3’-cNMP代谢所涉及的核酸和蛋白质的分子水平细节， 以及下游的细胞表型。拟议的研究还将扩大对以下方面的认识 由于2‘，3’-cNMP是信使核糖核酸降解的产物，因此细菌内的细胞信使核糖核酸的衰变途径。 这项工作预计将产生以下预期结果。首先，它将识别蛋白质 负责体内2‘，3’-cNMP的产生和降解，以及2‘，3’-cNMP结合蛋白 可能控制下游的表型。这些研究还将突出2‘，3’-cNMP在体内的分布 细菌王国，并扩大了我们对原核生物内信使核糖核酸衰退的理解。第二， 拟议的工作将确定改变一系列细菌中2‘，3’-cNMP水平的影响，包括变化 基因表达、表型和关键代谢途径，包括核苷酸/核苷代谢。 第三，阐明改变2‘，3’-cNMP水平和受2‘，3’-cNMP控制的表型的条件将 强调它们在细菌对细胞应激反应中的作用，特别是在细胞增殖方面 和生物膜的形成，并阐明了胁迫对mRNA衰退的额外影响。拟议的工作将 通过剖析2‘，3’-cNMP在细菌中的细胞作用，产生重要的积极影响，这将 突出原核生物中的新途径，并在未来可能被工程控制 细菌繁殖。

Abstract Novel intracellular small molecules, 2’,3’-cyclic nucleotide monophosphates (2’,3’-cNMPs), have recently been discovered within both prokaryotes and eukaryotes. Within plants and mammals, wounding has been found to cause increased levels of 2’,3’-cNMPs. Preliminary studies in bacteria suggest that 2’,3’-cNMPs also are produced in response to cellular stress and that 2’,3’-cNMP levels affect bacterial phenotypes, such as biofilm formation and motility, and expression of numerous genes. The long-term goal of this research is to understand the roles of 2’,3’-cNMPs in controlling prokaryotic signaling pathways and to utilize this knowledge to design small molecules to modulate cellular phenotypes. This goal will be addressed by investigating the cellular components involved in 2’,3’-cNMP metabolism, identifying 2’,3’-cNMP sensors, and determining the effects of 2’,3’-cNMPs on downstream pathways in a wide range of bacteria. The proposed work outlines an innovative research plan to probe a novel cellular stress-sensing mechanism and will provide molecular level details about the nucleic acids and proteins involved in 2’,3’-cNMP metabolism, as well as the downstream cellular phenotypes. The proposed studies also will expand knowledge of cellular mRNA decay pathways within bacteria because 2’,3’-cNMPs are products of mRNA degradation. This work is anticipated to yield the following expected outcomes. First, it will identify the proteins responsible for 2’,3’-cNMP production and degradation in vivo, as well as 2’,3’-cNMP binding proteins that may control downstream phenotypes. These studies also will highlight the distribution of 2’,3’-cNMPs within the bacterial kingdom and extend our understanding of mRNA decay within prokaryotes. Second, the proposed work will identify the effects of altering 2’,3’-cNMP levels in an array of bacteria, including changes in gene expression, phenotypes, and key metabolic pathways, including nucleotide/nucleoside metabolism. Third, elucidating conditions that alter 2’,3’-cNMP levels and phenotypes controlled by 2’,3’-cNMPs will highlight their role in bacterial responses to cellular stress, particularly with regards to cellular proliferation and biofilm formation, and illuminate additional effects of stress on mRNA decay. The proposed work will have an important positive impact by dissecting the cellular roles of 2’,3’-cNMPs within bacteria, which will highlight novel pathways within prokaryotes and, in the future, potentially can be engineered to control bacterial proliferation.