Modeling mechanisms of Shigella pathogenesis in yeast

病原菌的基因组正在迅速测序。一个主要的挑战是设计 筛选细菌基因组以鉴定致病蛋白质的高效、灵敏和特异性测定 并确定其在发病机制中的具体作用。虽然酵母不能作为生理 人类感染模型，我们的实验室和其他人最近建立了酵母菌 酿酒酵母作为一个强大的模型系统，研究细菌蛋白质的目标，潜在的保守 真核宿主细胞过程。本提案中提出的初步证据表明， 志贺氏菌蛋白赋予的毒性酵母表型是一种敏感和特异的筛选， 靶向宿主细胞过程的蛋白质。考虑到它相对较小的基因组，遗传上的易处理性，嗯- 开发后基因组工具，保护许多基本的细胞过程，以及丰富的 可用的系统数据S.酿酒酵母是多学科系统的理想模式生物， 生物学研究。为响应PA-02 - 011 "生物工程研究赠款"，我们建议： 开发和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、细胞生物学和新的生物信息学软件开发以鉴定宿主细胞 三种志贺氏菌蛋白IpgB，OspCI和OspF靶向的过程。证据表明 这些蛋白质中的每一种都在感染过程中直接递送到宿主细胞中，但对它们的作用知之甚少。 其内部的功能。我们假设，在这项提案中描述的全基因组筛选将 导致这些蛋白质中的每一种在发病机理中的分子作用的表征。 这项提议的实验集中在可遗传操纵的志贺氏菌蛋白质上， 可以相对容易地测试疾病生理模型中的假设。然而，一旦优化， 这种多学科的方法应该适用于研究任何微生物病原体， 细胞内宿主细胞过程，特别是对生长有危险或难以 像分枝杆菌和衣原体一样进行基因改造 这项工作很重要，与公共卫生问题有关，因为调查机制 细菌病原体用来引起疾病，我们将获得信息，这将有助于开发新的 抗生素来治疗这些感染。此外，我们有兴趣开发一种新的， 这是一种有效的方法来研究细菌病原体，这些病原体是危险的，难以以其他方式研究。

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.