嗜吞噬无形体四型分泌系统(T4SS)在调控宿主细胞凋亡和促胞内生存过程中发挥重要作用。已有研究表明T4SS效应蛋白Ats-1抑制线粒体介导细胞凋亡，但具体机制尚未阐明。本项目以效应蛋白Ats-1为切入点，利用转染、iTRAQ和生物信息学等技术，分析受Ats-1调控的宿主蛋白差异表达谱，筛选与细胞凋亡相关蛋白；利用酵母双杂交、酵母共转化和免疫沉淀等筛选验证与LGAS1、TIMP1互作的宿主细胞靶蛋白；基于上述实验结果，进行生物信息分析，寻找受Ats-1调控的细胞凋亡相关关键蛋白，利用RNA干扰、细菌侵染、实时荧光定量PCR、蛋白免疫印记及流式细胞术等技术，验证并揭示这些细胞凋亡相关关键蛋白与细胞凋亡信号通路关系，分析阐明效应蛋白Ats-1及其互作蛋白LGAS1、TIMP1参与的细胞凋亡信号通路及调控的分子机制。这将为开发新药物治疗靶标、新疫苗及人兽共患无无形体病防控提供科学依据。

Type Fourth Secretion System(T4SS) from Anaplasma phagocytophilum plays pivotal roles in regulating host cells apoptosis and promoting pathogen intracellular propagation and survives. The recent research demonstrated that effector Ats-1 inhibited mitochondrial-mediating apoptotic pathway, but the specific mechanism remains unclear. In this study, we will analyze and verificate host protein difference expression profile and apoptosis related proteins regulated by Ats-1 through transfection, isobaric tags for relative and absolute quantitation (iTRAQ) and bioinformatics; Screening host targets interaction with LGAS1、TIMP1 by yeast two hybride,yeast co-transformation and IP;The relations between the apoptosis related difference proteins and apoptosis pathway would further analyzed through RNA inference, pathogen infection, quantitative real-time PCR, Western Blot flow cytometry and so on. The molecular mechanism and singnal pathway of apoptosis regulated by Ats-1, and LGAS1,TIMP1 interaction with Ats-1 was intensive illuminate. These works will be useful for us find novel therapeutic targets, develop new preventive vaccines and take some clues for HGA control.