Acetylation of virulence factor regulatory proteins in V. cholerae

霍乱弧菌毒力因子调节蛋白的乙酰化

• 批准号: 9896576
• 负责人: Claudia C Hase
• 金额: $ 17.86万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-12 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9896576
• 关键词: Acetyl Coenzyme A; Acetylation; Acetyltransferase; Affect; Amino Acids; Animal Model; Bacteria; Biochemical; Biological Process; Carbon; Cellular Metabolic Process; Citric Acid Cycle; Collaborations; Complex; DNA; Data; Diarrhea; Digestion; Disease; Environment; Enzymes; Exposure to; Food; Future; Gastrointestinal tract structure; Gelshift Analysis; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Goals; Growth; Human; Infection; Intervention; Knowledge; Lead; Link; Lysine; Mass Spectrum Analysis; Membrane; Metabolic; Metabolic Pathway; Metabolism; Modernization; Modification; Molecular; Mutation; Nutrient; Organism; Pathogenesis; Pathogenicity; Pathway interactions; Pattern; Play; Post-Translational Protein Processing; Process; Production; Protein Acetylation; Proteins; Regulation; Reporting; Research; Research Design; Respiration; Role; Signal Transduction; Site; Small Intestines; Source; Testing; Time; Variant; Vibrio cholerae; Virulence; Virulence Factors; Work; absorption; antimicrobial; bacterial metabolism; base; flexibility; genetic regulatory protein; human pathogen; in vivo; insight; microorganism; mutant; novel; pathogen; pathogenic bacteria; response; transcription factor; treatment strategy

Abstract The metabolism of pathogenic microorganisms has to be flexible and the efficient replication of pathogenic bacteria in the host depends on an active adaptation process. Vibrio cholerae is exposed to a variety of environments within the human host and eventually has to adapt to the nutrients available at its preferred infection site, the small intestine. The small intestine is the part of the gastrointestinal tract where the majority of digestion and absorption of food takes place, thus providing a variety of nutrients for the bacteria. As V. cholerae start to cause the profuse diarrhea that is the hallmark of the disease, the bacteria have to once again adapt to the resulting changes in nutrient availability to continue the infection process. It can therefore be expected that the metabolic status in V. cholerae changes substantially during the course of infection and that changes in metabolism are coordinated with virulence gene expression. Although a link between the metabolic status and virulence factor expression has been well described in several pathogens, it has not been investigated in detail in V. cholerae. Our preliminary results suggest that different availability of certain carbon sources that have relevance to in vivo conditions affect virulence gene expression in V. cholerae. Moreover, we found that mutations in membrane respiration as well as central metabolism genes, such as enzymes involved in the TCA cycle, affect virulence factor expression in V. cholerae. Based on our observations with defined mutants in the PTA-ACK pathway we concluded that acetyl-CoA (acCoA), but not acetyl-phosphate, plays a role in this signal transduction. We hypothesize that this link between metabolism and virulence gene regulation in V. cholerae occurs via post-translational protein modification, specifically by lysine acetylation of key virulence regulatory proteins. Importantly, a recent study of the global acetylome of V. cholerae revealed lysine acetylation of several important virulence regulatory proteins, including TcpP, AphB, and PhoB. By combining modern mass spectrometry analyses with genetic and biochemical analyses we expect to better understand the mechanism by which acCoA affects virulence factor transcription. We will also examine the roles of putative V. cholerae acetyltransferase genes in virulence factor modification as well as determine additional overall protein acetylation patters (acetylomes). Thus, our study has the potential to, for the first time, demonstrate a role of lysine acetylation in V. cholerae virulence factor regulation and will further establish the importance of central metabolism in the pathogenesis of this organism. Overall, we expect to gain insights into the metabolic pathways of this organism per se and its relation to virulence as an important aspect of host- pathogen interaction. Ultimately, this information could lead to novel intervention strategies aimed at modulating the interplay of central metabolites and virulence factor expression, not only for V. cholerae but potentially other pathogens.

抽象的 致病微生物的代谢必须是灵活的，并且有效复制 宿主中的致病细菌取决于主动适应过程。 Vibrio Cholerae暴露于 人类宿主中的各种环境，最终必须适应其可用的营养 首选感染部位，小肠。小肠是胃肠道的一部分 大多数消化和食物吸收发生，因此为细菌提供了多种营养。 由于霍乱谷开始引起疾病标志的大量腹泻，因此细菌必须一次 再次适应养分可用性的结果变化，以继续感染过程。因此可以是 预计霍乱弧菌的代谢状态在感染过程中发生了很大变化，并且 代谢的变化与毒力基因表达协调。虽然代谢之间的联系 状态和毒力因子表达在几种病原体中已经很好地描述了 在V.霍乱中进行了详细研究。我们的初步结果表明某些碳的可用性不同 与体内条件相关的来源会影响霍乱弧菌中的毒力基因表达。而且， 我们发现膜呼吸中的突变以及中央代谢基因，例如酶 参与TCA周期，影响霍乱弧菌中的毒力因子表达。根据我们对 在PTA-ACK途径中定义的突变体我们得出结论，乙酰基-COA（ACCOA），但不是乙酰磷酸乙酰乙醇 在此信号转导中起作用。我们假设代谢与毒力基因之间的这种联系 霍乱弧菌中的调节是通过翻译后蛋白质修饰发生的，特别是通过赖氨酸乙酰化的 关键的毒力调节蛋白。重要的是，对霍乱弧菌的全球乙酰基组的最新研究表明 包括TCPP，APHB和PHOB在内的几种重要毒力调节蛋白的赖氨酸乙酰化。经过 将现代质谱分析与我们期望更好的遗传和生化分析相结合 了解ACCOA影响毒力因子转录的机制。我们还将检查 推定的V.霍乱乙酰基转移酶基因在毒力因子修饰中的作用，并确定 额外的总体蛋白质乙酰化模式（乙酰基组）。因此，我们的研究有潜力，第一个 时间，证明赖氨酸乙酰化在霍乱弧菌毒力因子调节中的作用，并将进一步建立 中央代谢在这种生物体的发病机理中的重要性。总体而言，我们希望获得见解 进入这种生物本身的代谢途径及其与毒力的关系，作为宿主的重要方面 病原体相互作用。最终，这些信息可能导致针对的新颖干预策略 调节中央代谢产物和毒力因子表达的相互作用，不仅用于霍乱弧菌 潜在的其他病原体。