Global regulators converge to orchestrate metabolism, biofilm, and pathogenesis

全球监管机构齐心协力协调代谢、生物膜和发病机制

• 批准号: 10380787
• 负责人: PAULA I WATNICK
• 金额: $ 59.17万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10380787
• 关键词: Attenuated; Bacteria; Binding; Binding Proteins; Caring; Cell membrane; Cells; ChIP-seq; Childhood; Cholera; Chromosomes; Climate; Complex; Cues; Cyclic AMP Receptor Protein; Cytoplasm; Development; Diarrhea; Diet; Diet Modification; Dietary Intervention; Disease; Dissociation; Electrostatics; Environment; Enzymes; Funding; Gene Activation; Gene Expression; Genetic Transcription; Glucose; Goals; Grant; Growth; In Vitro; Incidence; Integral Membrane Protein; Internet; Intestines; Laboratories; Location; Lysine; Medical; Membrane; Membrane Lipids; Membrane Proteins; Metabolic; Metabolic Control; Metabolic Pathway; Metabolism; Microbial Biofilms; Modeling; Modification; Morbidity - disease rate; N-terminal; Nature; Nutrient; Nutritional; Pathogenesis; Pathogenicity; Post-Translational Protein Processing; Process; Protein translocation; Proteins; Regulation; Regulon; Research; Resource-limited setting; Role; Signal Transduction; Surface; Transcriptional Regulation; Vibrio cholerae; Virulence; Virulence Factors; Work; base; design; diarrheal disease; dietary; follow-up; genetic regulatory protein; gut colonization; in vivo; mortality; pandemic disease; pathogen; pathogenic bacteria; prevent; promoter; response; transcription factor; uptake

Project Summary Diarrheal disease is a leading cause of morbidity and mortality in resource-poor areas. In order to colonize the intestine and cause disease, successful bacterial pathogens must sense and respond to intestinal signals by altering both metabolism and virulence factor expression. We hypothesize that by understanding the critical intestinal signals and the bacterial regulatory networks they activate, we can devise simple dietary alterations that prevent or mitigate morbidity and mortality. We focus on Vibrio cholerae, the agent of the severe diarrheal disease cholera. Because the metabolic pathways we study are highly conserved, these findings also serve as a paradigm for other bacteria that cause diarrhea. The goal of this work is to elucidate the complex and highly conserved regulatory network that is activated when V. cholerae enters the intestinal environment. In the first 4-year funding period of this grant, we explored the role of a global regulator of metabolism and virulence known as glucose-specific Enzyme llA. We showed that this regulator is membrane-associated through an N-terminal amphipathic helix and that membrane association is critical for its interaction with integral membrane protein partners that it regulates. Based on this work, we hypothesize that the inner membrane of the bacterial cell may act as a platform for regulatory proteins that sense and respond to nutritional signals in the intestinal environment. During the previous funding period, we discovered that the subcellular location of the global transcription factor, the cAMP receptor protein (CRP), is regulated in response to environmental conditions. In the current funding period, we propose to follow up on these observations by investigating regulation of CRP subcellular localization, the mechanism by which subcellular localization alters CRP activation of gene transcription, and the functional significance of CRP subcellular localization for V. cholerae virulence. We hypothesize that if the subcellular localization of CRP can be controlled via host diet, it may be possible to reduce pathogen virulence and thereby the morbidity and mortality caused by cholera.

项目摘要 腹泻病是资源贫乏地区发病率和死亡率的主要原因。为了殖民 肠道并引起疾病，成功的细菌病原体必须感知并通过 改变新陈代谢和毒力因子的表达。我们通过理解关键来假设 肠道信号和它们激活的细菌调节网络，我们可以设计简单的饮食改变 这可以防止或减轻发病率和死亡率。 我们专注于严重腹泻病霍乱的毒蛇霍乱。因为代谢 我们研究的途径是高度保守的，这些发现也可以作为其他引起的细菌的范例 腹泻。这项工作的目的是阐明复杂且高度保守的监管网络 当V.霍乱进入肠道环境时被激活。 在这笔赠款的第一个4年资金期间，我们探讨了全球代谢监管者和 毒力称为葡萄糖特异性酶LLA。我们表明该调节剂与膜相关 通过N末端两亲性螺旋，该膜关联对于与其与之相互作用至关重要 它调节的整体膜蛋白伙伴。基于这项工作，我们假设内在 细菌细胞的膜可以充当调节蛋白的平台，并回应 肠道环境中的营养信号。 在上一个资金期间，我们发现全局转录的亚细胞位置 因素，cAMP受体蛋白（CRP）受到环境条件的响应。在电流中 我们建议通过研究CRP亚细胞的调节来跟进这些观察结果 定位，亚细胞定位改变基因转录的CRP激活的机制， CRP亚细胞定位对霍乱弧菌毒力的功能意义。我们假设如果 可以通过宿主饮食控制CRP的亚细胞定位，可能会降低病原体的毒力 从而由霍乱引起的发病率和死亡率。