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: 9249525
• 负责人: Sara Marie Raser Palmer
• 金额: $ 24.9万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9249525
• 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; Data Analyses; Dental caries; Disease; Endocarditis; Enrollment; Etiology; Family; Gene Cluster; Gene Expression Regulation; Genes; Genetic; Genetic Transformation; Genome; Genomics; Goals; Growth; Individual; Infection; Investigation; Knowledge; Life; Membrane; Membrane Proteins; Mentors; Microbial Biofilms; Mitochondria; Modeling; Molecular; Organism; Oxidative Stress; Pancreatic ribonuclease; Pathogenesis; Pathway interactions; Phase; Phenotype; Play; Post-Transcriptional Regulation; Production; Property; Proteins; Proton-Translocating ATPases; RNA; RNA analysis; RNA-Binding Proteins; Rattus; Regulation; Research; Role; Scientist; Sequence Analysis; Small RNA; Streptococcus mutans; Stress; Stroke; Techniques; Training; Universities; Virulence; Virulence Factors; Work; acid stress; career; career development; cariogenic bacteria; cell envelope; combat; deep sequencing; dental agent; design; differential expression; effective therapy; experience; experimental study; gene function; genome analysis; genome-wide; insight; lactic acid bacteria; member; membrane biogenesis; mutant; novel; pan-genome; paralogous gene; post-doctoral training; pre-doctoral; programs; protein expression; public health relevance; response; stress tolerance; tool; transcriptome; transcriptome sequencing; 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）的主要病因，但这种细菌也会引起心内膜炎，并与心血管疾病和中风密切相关。绝大多数使变形链球菌致病的因子位于细胞质膜或分泌于细胞外。变形链球菌包膜的正常生物发生需要两个同源的普遍保守的YidC/Oxa/Alb3家族的YidC蛋白。在大鼠龋齿模型中，删除任意一个yidC基因导致突变链球菌毒力降低，而删除两个基因则是致命的。此外，在yidC突变体中，胁迫耐受性、H+- atp酶活性和生物膜形成均下降，且与yidC1相比，缺失yidC2的影响更为严重。此外，有强有力的证据表明，yidC1和yidC2在乳酸菌中受到不同的调节。在最近的合作中，我们对57个临床分离的变形链球菌进行了基因组测序和分析，表明泛基因组和核心基因组分别包含约3300和1400个基因。本研究还预测了S. mutans中的一些小rna，通过rna深度测序，我们确定了一些sRNAs的差异表达。作为申请人博士后培训的一部分，对15个测序菌株进行了广泛的表型表征，并证明在基因含量和毒力潜力方面存在实质性的多样性