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.

摘要 病原微生物的代谢必须是灵活的， 病原菌在宿主体内的生存依赖于一个积极的适应过程。霍乱弧菌暴露于 在人类宿主的各种环境中，最终必须适应其体内的营养物质。 感染的首选部位是小肠小肠是胃肠道的一部分， 食物的大部分消化和吸收发生，从而为细菌提供各种营养。 随着霍乱弧菌开始引起大量腹泻，这是疾病的标志，细菌必须一旦 再次适应由此产生的营养可用性变化以继续感染过程。因此可以 预期霍乱弧菌的代谢状态在感染过程中发生了显著变化， 代谢的变化与毒力基因的表达相协调。尽管新陈代谢和 状态和毒力因子表达已在几种病原体中得到了很好的描述，但还没有 详细研究了霍乱弧菌。我们的初步结果表明，不同的碳的可用性， 与体内条件相关的来源影响霍乱弧菌中的毒力基因表达。此外，委员会认为， 我们发现，细胞膜呼吸和中枢代谢基因（如酶）的突变， 参与TCA循环，影响霍乱弧菌中毒力因子的表达。根据我们对 在PTA-ACK途径中定义的突变体，我们得出结论，乙酰辅酶A（acCoA），而不是乙酰磷酸， 在信号传导中起着重要作用。我们假设代谢和毒力基因之间的这种联系 霍乱弧菌中的调节通过翻译后蛋白质修饰发生，特别是通过 关键毒力调节蛋白。重要的是，最近一项关于霍乱弧菌全局乙酰组的研究显示， 赖氨酸乙酰化的几个重要的毒力调节蛋白，包括TcpP，AphB，和PhoB。通过 将现代质谱分析与遗传和生物化学分析相结合， 了解acCoA影响毒力因子转录的机制。我们亦会研究 假定的霍乱弧菌乙酰转移酶基因在毒力因子修饰中的作用以及确定 额外的整体蛋白质乙酰化模式（乙酰体）。因此，我们的研究有可能，首先， 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 中枢代谢在该生物体发病机制中的重要性。总的来说，我们希望获得 了解这种生物本身的代谢途径及其与毒力的关系，毒力是宿主的一个重要方面， 病原体相互作用最终，这些信息可能会导致新的干预策略， 调节中心代谢产物和毒力因子表达的相互作用，不仅对于霍乱弧菌， 可能是其他病原体。