pORF5与Pin1相互作用对沙眼衣原体生长发育的影响及其机制研究

沙眼衣原体（Ct）的生长发育依赖于其分泌蛋白与宿主细胞之间复杂的cross-talk。pORF5是申请者首次发现由Ct质粒基因编码并主要定位于宿主细胞胞浆的一种分泌蛋白。前期研究采用蛋白质组学技术发现pORF5上调宿主肽基脯氨酰顺反异构酶Pin1的表达，免疫共沉淀证实二者为相互作用蛋白，但两者相互作用对Ct生长发育的影响及机制尚不明确。本项目在此基础上，首先采用基因转染、抗体阻断和siRNA等从细胞和动物水平上研究pORF5与Pin1相互作用对Ct生长发育的影响，然后采用串联亲和纯化结合质谱技术、生物信息学技术等筛选pORF5功能相关的Pin1蛋白复合物，构建蛋白相互作用网络，确认与Ct生长发育相关的关键蛋白和KEGG/BIOCARTA信号通路并探讨其生物学功能，揭示Ct通过pORF5与Pin1相互作用促进Ct生长发育的网络调控机制。本研究能丰富对Ct致病机制的认识，也为其防治提供依据。

Growth of Chlamydia trachomatis(Ct) depends on the complex cross-talk between the Chlamydial secreted proteins and host cell. Our previous work has demonstrated that pORF5 encoded by chlamydial plasmid was a secreted protein and mainly located in cytoplasm of Ct-infected host cells for the first time. Pin1, a peptidyl-prolyl isomerase was screened to be up-regulated in pORF5-tranfected cells by using proteomic technique, we further confirmed that the two proteins interacted with each other by co-immunoprecipitation. However, the effect and mechanism of interaction between pORF5 and Pin1 proteins on Ct growth are still kept unkown. Based on our fingdings, this project first intends to apply gene transfection, antibody blocking and siRNA to identify the effect of interaction between pORF5 and Pin1 proteins on Ct growth in cell and animal models. Then tandem affinity purification combined with mass spectrometry and bioinformatic technique will be used to screen Pin1-interacting proteins and map the protein interaction network in pORF5-transfected cells, and also to elucidate the roles of key target proteins and KEGG/BIOCARTA signal pathways for Ct growth. This work will reveal the molecular mechanism and regulatory network responsible for Chlamydial growth mediated by interaction between pORF5 and Pin1, which not only enriches the understanding of Ct pathogenesis, but also provides a basis for its prevention and treatment.