Modeling mechanisms of Shigella pathogenesis in yeast

酵母志贺氏菌发病机制的建模机制

基本信息

批准号：
8093565
负责人：
CAMMIE LESSER
金额：
$ 10万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-19 至 2011-12-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8093565
关键词：
Animal Model Bacterial Genome Bacterial Proteins Bioinformatics Biological Assay Biological Models Biomedical Engineering Cell physiology Cells Cellular biology Chlamydia Co-Immunoprecipitations Computer software Coupled Data Disease Dominant-Negative Mutation Epithelial Cells Eukaryota Gene Expression Genetic Genome Genomics Genus Mycobacterium Human Immunofluorescence Microscopy Indirect Immunofluorescence Individual Infection Investigation Laboratories Liquid Chromatography Mediating Modeling Molecular Monitor Pathogenesis Pathway interactions Pattern Phenotype Plaque Assay Protein Microchips Proteins Proteomics Public Health RNA Interference Research Personnel Research Project Grants Robotics Role Saccharomyces cerevisiae Shigella System Systems Biology Techniques Technology Testing Time Transmission Electron Microscopy Work Yeasts antimicrobial data mining design genome wide association study genome-wide interdisciplinary approach interest microbial multidisciplinary novel pathogen pathogenic bacteria physiologic model programs protein complex research study response software development tandem mass spectrometry tool

项目摘要

The genomes of pathogenic bacteria are being rapidly sequenced. A major challenge is to devise efficient, sensitive, and specific assays to screen bacterial genomes to identify pathogenic proteins and determine their specific roles in pathogenesis. Although yeast cannot serve as a physiologic model of human infection, our laboratory and others have recently established Saccharomyces cerevisiae as a powerful model system to study bacterial proteins that target potentially conserved eukaryotic host cell processes. Preliminary evidence presented in this proposal demonstrates that toxic yeast phenotypes conferred by Shigella proteins are a sensitive and specific screen for proteins that target host cell processes. Given its relatively small genome, genetic tractability, well- developed post-genomic tools, conservation of many basic cellular processes, and the wealth of available systematic data S. cerevisiae is an ideal model organism for multidisciplinary systems- biology studies. In response to PA-02-011, "Bioengineering Research Grants," we propose to develop and validate a multidisciplinary, integrative, systems approach in yeast involving genomics, proteomics, cell biology and novel bioinformatics software development to identify host cell processes targeted by three Shigella proteins, IpgB, OspCI and OspF. Evidence suggests that each of these proteins is delivered directly into host cells during infection, but little is known about their functions within. We hypothesize that the genome-wide screens described in this proposal will result in the characterization of the molecular roles in pathogenesis of each of these proteins. Experiments in this proposal focus on proteins from the genetically manipulable Shigella so that we can relatively easily test hypotheses in physiologic models of disease. However, once optimized, this multidisciplinary approach should be applicable to study any microbial pathogen that targets intracellular host cell processes, especially pathogens that are dangerous to grow or difficult to genetically manipulate like Mycobacterium and Chlamydia. This work is important is relevant to public health issues since by investigations mechanisms that bacterial pathogens used to cause disease, we will gain information that will help develop new antimicrobials to treat these infections. Furthermore, we are interested in developing a new and efficient way to study bacterial pathogens that are dangerous and difficult to study in other ways.

致病细菌的基因组正在快速测序。一个主要的挑战是设计高效、灵敏且特异的检测方法可筛选细菌基因组以识别致病蛋白并确定它们在发病机制中的具体作用。虽然酵母不能起到生理作用我们的实验室和其他人最近建立了人类感染模型酿酒酵母作为一个强大的模型系统来研究潜在保守的细菌蛋白质真核宿主细胞过程。该提案中提供的初步证据表明志贺氏菌蛋白赋予的有毒酵母表型是一种敏感且特异的筛选方法靶向宿主细胞过程的蛋白质。鉴于其基因组相对较小、遗传易处理性，开发了后基因组工具，保护了许多基本的细胞过程，以及丰富的现有的系统数据酿酒酵母是多学科系统的理想模式生物- 生物学研究。为了响应 PA-02-011“生物工程研究补助金”，我们建议开发并验证涉及基因组学的酵母多学科、综合系统方法，蛋白质组学、细胞生物学和新型生物信息学软件开发，用于识别宿主细胞三种志贺氏菌蛋白 IpgB、OspCI 和 OspF 所针对的过程。有证据表明这些蛋白质中的每一种都在感染过程中直接传递到宿主细胞中，但人们对它们知之甚少他们的职能在里面。我们假设本提案中描述的全基因组筛选将导致这些蛋白质在发病机制中的分子作用的表征。该提案中的实验重点关注来自可基因操纵志贺氏菌的蛋白质，以便我们可以相对容易地检验疾病生理模型中的假设。然而，一旦优化后，这种多学科方法应该适用于研究任何针对目标的微生物病原体细胞内宿主细胞过程，尤其是生长危险或难以生长的病原体像分枝杆菌和衣原体一样进行基因操纵。这项工作很重要，与公共卫生问题相关，因为通过调查机制细菌病原体用于引起疾病，我们将获得有助于开发新的信息抗生素来治疗这些感染。此外，我们有兴趣开发一种新的和研究危险且难以通过其他方式研究的细菌病原体的有效方法。