Chlamydia trachomatis Secreted Effector Proteins: Infection Properties and Identification of Host Targets

项目摘要 沙眼衣原体是最常见的性传播疾病的病原体。 在美国。疾病在女性中最为严重，可能对生殖健康造成有害影响， 比如不孕不育。在非免疫细胞内建立细胞内生态位，如粘膜上皮， 足以引发免疫病理学和疾病后遗症。同时缺乏TMEA和TmeB的沙眼衣原体 III分泌系统效应器，在组织培养中产生较少的感染性后代，并显著 降低小鼠的传染性。以前利用荧光识别它们各自功能的尝试-- 已报道的等位基因交换突变(FRAEM)，但由于盒诱导而导致表型不明 极地效应。最近，我开发了FLAEM(FLAEM)，以逆转 并产生第一个无标记沙眼衣原体缺失突变体。我 假设TMEA和TmeB在功能上相关，并且对于建立包含关系很重要，因为 它们是共转录的，共享一个共同的伴侣，同时分泌到宿主细胞中。这个 拟议的工作将利用新的无标记沙眼衣原体缺失突变体来确定何时 在衣原体发育过程中，TMEA和TmeB起重要作用，并鉴定它们的真核宿主细胞 目标。这项研究的结果将直接有助于我们对基础沙眼衣原体的理解。 生物学和他们如何使用分泌的效应器来建立细胞内，这将识别潜在的药物靶点 以防止感染传播到全球数百万人。

Project Summary Chlamydia trachomatis is the causative agent of most commonly reported sexually transmitted disease in the United States. Disease is most severe in women and can cause detrimental effects to reproductive health, such as infertility. Establishment of an intracellular niche within non-immune cells, such as mucosal epithelium, is sufficient to drive immunopathology and disease sequela. C. trachomatis lacking both TmeA and TmeB, type III secretion system effectors, generate fewer infectious progeny in tissue culture and have significantly decreased infectivity in mice. Previous attempts to identify their individual functions utilized fluorescence- reported allelic exchange mutagenesis (FRAEM), but resulted in unclear phenotypes due to cassette-induced polar effects. Recently, I have developed floxed-cassette allelic exchange mutagenesis (FLAEM), to reverse cassette-induced polar effects and generate the first ever markerless C. trachomatis deletion mutant. I hypothesize that TmeA and TmeB are functionally related and important for establishing the inclusion, because they are co-transcribed, share a common chaperone, and are secreted into the host cell at the same time. The proposed work will utilize the novel marker-less C. trachomatis deletion mutant to determine when during chlamydial development TmeA and TmeB are important, and identify their eukaryotic host cell targets. The outcomes of this research will directly contribute to our understanding of basic C. trachomatis biology and how they use secreted effectors to establish an intracellular, which will identify potential drug targets to prevent the spread of infection to millions world-wide.