Genetics and Genomics of the yidC paralogs in Streptococcus mutans

变形链球菌 yidC 旁系同源物的遗传学和基因组学

• 批准号: 8618281
• 负责人: Sara Marie Raser Palmer
• 金额: $ 10.8万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-13 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8618281
• 关键词: Adherence; Advisory Committees; Affect; Area; Atherosclerosis; Award; Bacteria; Bacterial Infections; Biogenesis; Bioinformatics; Carbohydrates; Cardiovascular Diseases; Cell membrane; Cell physiology; Cell surface; Cells; Chloroplasts; Clinical; Competence; Counseling; Data; Dental caries; Disease; Endocarditis; Enrollment; Equilibrium; Family; Gene Cluster; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Transformation; Genome; Genomics; Goals; Growth; Individual; Infection; Knowledge; Life; Membrane; Membrane Proteins; Mentors; Microbial Biofilms; Mitochondria; Modeling; Molecular; Organism; Oxidative Stress; Pathogenesis; Pathway interactions; Phase; Phenotype; Play; Post-Transcriptional Regulation; Production; Property; Proteins; Proton-Translocating ATPases; RNA; RNA Sequence Analysis; RNA Sequences; RNA-Binding Proteins; Rattus; Regulation; Research; Ribonucleases; Role; Scientist; Sequence Analysis; Small RNA; Streptococcus mutans; Stress; Stroke; Techniques; Training; Universities; Virulence; Virulence Factors; Work; acid stress; base; career; career development; cariogenic bacteria; cell envelope; combat; deep sequencing; design; effective therapy; experience; gene function; insight; lactic acid bacteria; member; membrane biogenesis; mutant; novel; paralogous gene; post-doctoral training; pre-doctoral; programs; protein expression; public health relevance; research study; response; stress tolerance; tool; transcriptomics

DESCRIPTION (provided by applicant): Streptococcus mutans is the primary etiological agent of dental caries (cavities), but the bacterium also causes endocarditis and is strongly associated with cardiovascular disease and stroke. The vast majority of the factors that allow S. mutans to cause disease are located in the cytoplasmic membrane or are secreted outside of the cell. Proper biogenesis of the cell envelope of S. mutans requires two paralogous YidC proteins of the universally conserved YidC/Oxa/Alb3 family. Deletion of either yidC gene results in diminished virulence of S. mutans in a rat caries model, while deletion of both genes is lethal. Moreover, stress tolerance, H+-ATPase activity, and biofilm formation are decreased in yidC mutants, with more severe effects observed upon deletion of yidC2 compared to yidC1. Additionally, there is strong evidence that yidC1 and yidC2 are differentially regulated in lactic acid bacteria. In a recent collaborative effort, we sequenced and analyzed the genomes of 57 clinical isolates of S. mutans, demonstrating that the pan-, and core-genomes contain approximately 3300 and 1400 genes, respectively. This study also predicted a number of small RNAs in S. mutans, and through RNA-deep sequencing we determined that some of the sRNAs are differentially expressed. As part of the postdoctoral training of the applicant, an extensive phenotypic characterization was performed on 15 of the sequenced strains, and it was demonstrated that there is substantial diversity in gene content and virulence potential within the species S. mutans. The major scientific goals of this proposal are to dissect the molecular basis for regulation of yidC1 and yidC2, and to provide a comprehensive understanding of the roles of these proteins in stress adaptation and cellular physiology across the species S. mutans. To accomplish these goals, the following Specific Aims are delineated: Aim 1. Determine the mechanisms of regulation of yidC1 and yidC2 in response to growth domain, and to environmental influences that are known to induce membrane stress and to affect persistence and pathogenesis by S. mutans. Aim 2. Investigate the role of small RNAs, a ribonuclease, and an RNA binding protein encoded within the yidC gene clusters in the regulation of yidC expression and function of the YidC proteins Aim 3. Determine how deletion of yidC1 or yidC2 affects the transcriptomes of phenotypically and genetically-diverse isolates of S. mutans using RNA-deep sequencing. The completion of this project will also allow the applicant to receive critical training in the areas of gene regulation and genome-scale analytical techniques, while obtaining the mentoring and career development counseling to facilitate her transition to an independent research career. The data generated will provide essential new information that can be used to design novel and more effective therapies to combat dental caries and other bacterial infections.

描述（由申请人提供）：变形链球菌是龋齿（蛀牙）的主要病原体，但该细菌也会引起心内膜炎，并与心血管疾病和中风密切相关。变形链球菌引起疾病的绝大多数因子位于细胞质膜中或分泌到细胞外。变形链球菌细胞包膜的正确生物发生需要普遍保守的 YidC/Oxa/Alb3 家族的两种旁系同源 YidC 蛋白。删除任一 yidC 基因都会导致大鼠龋齿模型中变形链球菌的毒力减弱，而删除这两个基因则是致命的。此外，yidC 突变体的胁迫耐受性、H+-ATP酶活性和生物膜形成均降低，与 yidC1 相比，删除 yidC2 后观察到更严重的影响。此外，有强有力的证据表明 yidC1 和 yidC2 在乳酸菌中受到差异性调节。在最近的一项合作中，我们对 57 个变形链球菌临床分离株的基因组进行了测序和分析，证明泛基因组和核心基因组分别包含大约 3300 个和 1400 个基因。这项研究还预测了变形链球菌中的一些小RNA，通过RNA深度测序，我们确定了一些sRNA存在差异表达。作为申请人博士后培训的一部分，对 15 个已测序菌株进行了广泛的表型表征，结果表明，该菌株的基因内容和毒力潜力存在很大的多样性。 种变形链球菌。该提案的主要科学目标是剖析 yidC1 和 yidC2 调节的分子基础，并全面了解这些蛋白质在变形链球菌物种的应激适应和细胞生理学中的作用。为了实现这些目标，描述了以下具体目标： 目标 1. 确定 yidC1 和 yidC2 响应生长域和环境影响的调节机制，已知环境影响会诱导膜应激并影响变形链球菌的持久性和发病机制。目标 2. 研究 yidC 基因簇内编码的小 RNA、核糖核酸酶和 RNA 结合蛋白在调节 yidC 表达和 YidC 蛋白功能中的作用。 目标 3. 使用 RNA 深度测序确定 yidC1 或 yidC2 的删除如何影响表型和遗传多样性的变形链球菌分离株的转录组。该项目的完成还将使申请人能够接受基因调控和基因组规模分析技术领域的关键培训，同时获得指导和职业发展咨询，以促进她向独立研究职业的过渡。生成的数据将提供重要的新信息，可用于设计新颖且更有效的疗法来对抗龋齿和其他细菌感染。