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: 8734378
• 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 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8734378
• 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。绝大多数的因素，使S。引起疾病的变异蛋白位于细胞质膜中或分泌到细胞外。S.变形杆菌需要普遍保守的YidC/Oxa/Alb 3家族的两种旁系同源YidC蛋白。任一yidC基因的缺失导致S.在大鼠龋齿模型中，两种基因的缺失都是致命的。此外，胁迫耐受性，H+-ATP酶的活性，和生物膜的形成减少yidC突变体，与更严重的影响后，观察到删除yidC 2相比yidC 1。此外，有强有力的证据表明yidC 1和yidC 2在乳酸菌中受到差异调节。在最近的一次合作中，我们对57株临床分离的沙门氏菌进行了基因组测序和分析。变异，表明泛基因组和核心基因组分别含有约3300和1400个基因。本研究还预测了S.通过RNA深度测序，我们确定了一些sRNA的差异表达。作为申请人博士后培训的一部分，对15种测序菌株进行了广泛的表型表征，并证明了菌株内的基因内容和毒力潜力存在显著的多样性。 种S.变异人该提案的主要科学目标是剖析yidC 1和yidC 2调控的分子基础，并全面了解这些蛋白质在整个物种S的应激适应和细胞生理学中的作用。变异体为了实现这些目标，制定了以下具体目标：目标1。确定yidC 1和yidC 2响应生长域的调节机制，以及已知诱导膜应激和影响S.变异人目标二。研究小RNA、核糖核酸酶和yidC基因簇内编码的RNA结合蛋白在调节yidC表达和YidC蛋白Aim 3功能中的作用。确定yidC 1或yidC 2的缺失如何影响表型和遗传多样性的S.使用RNA-deep sequencing。该项目的完成还将使申请人在基因调控和基因组规模分析技术领域接受关键培训，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。所产生的数据将提供重要的新信息，可用于设计新颖和更有效的治疗方法，以对抗龋齿和其他细菌感染。