Type III exported effectors of Chlamydia trachomatis

授予头衔：沙眼衣原体III型出口效应器 摘要： 人类病原体沙眼衣原体在美国是一个严重的问题，因为 它的流行以及急性和慢性疾病对健康和社会经济的综合影响 疾病。衣原体是专有的细胞内病原体，具有调节 寄主细胞在隔膜的寄生液泡中发挥作用。 尽管已经确定III型分泌系统(T3SS)代表一种机制 被用来调节关键的宿主细胞通路，对这一过程的总体理解 衣原体感染仍然很有限。在过去的筹资周期中，我们在以下方面取得了重大进展 了解指定为CT694和CT695的T3S效应器的作用。重要的是，我们还 利用新的遗传操作技术研究T3s对衣原体的影响 发病机制。也许我们最重要的贡献是开发了一种 荧光报告的等位基因交换突变(FRAEM)方法首次允许 时间，目标衣原体基因的缺失。我们已经使用此技术创建了空 Ct694和ct695的突变体，这些菌株在组织培养和 动物感染模型。我们建议阐明突变体的分子细节 表型表现明显。基因、生物化学、细胞生物学和 提出了基于动物的研究，将定义发育缺陷，确定宿主目标 对效应器功能的重要性(S)，并衡量效应器和靶蛋白在 促进衣原体致病。这项工作的完成将延长 从基因上操纵一种具有挑战性的病原体并导致对 衣原体引起的疾病。

Grant title: Type III exported effectors of Chlamydia trachomatis Abstract: The human pathogen Chlamydia trachomatis is a significant concern in the United States due to its prevalence and the combined health and socioeconomic impact of acute and chronic disease. Chlamydiae are obligate intracellular pathogens that possess the ability to modulate host-cell functions while sequestered within a membrane-bound parasitophorous vacuole. Although it is established that a type III secretion system (T3SS) represents one mechanism employed to modulate critical host cell pathways, the overall understanding of this process in Chlamydia remains limited. During the past funding cycle, we made significant progress in understanding the role of T3S effectors designated CT694 and CT695. Importantly, we also leveraged novel genetic manipulation techniques to investigate the impact of T3S in chlamydial pathogenesis. Perhaps our most important contribution was the development of a fluorescence-reported allelic exchange mutagenesis (FRAEM) approach to allow, for the first time, deletion of targeted chlamydial genes. We have used this technique to create null mutants for ct694 and ct695, and these strains are attenuated in both tissue-culture and animal-infection models. We propose to elucidate molecular details by which mutant phenotypes are manifested. A balanced combination of genetic, biochemical, cell-biology, and animal-based studies are proposed that will define developmental defects, identify host targets important for effector function(s), and gauge the overall role of effectors and target proteins in promoting Chlamydia pathogenesis. Completion of this work will extend the efficacy of genetically manipulating a challenging pathogen and lead to an enhanced understanding of Chlamydia-mediated disease.