烟曲霉COFILIN与人GOSR1蛋白相互作用并抑制HEK293T细胞的增殖

Aspergillus fumigatus COFILIN Interacts with Human GOSR1 and Inhibits HEK293T Cell Proliferation

COFILIN是真核生物中肌动蛋白的解聚因子,使细胞骨架产生活力.烟曲霉(Aspergillus fumigatus)COFILIN蛋白是其侵入机体的主要致病力因子,也是其逃逸宿主免疫系统防御的重要手段.为了探索烟曲霉COFILIN在感染宿主细胞过程中的作用机制,首先利用酵母双杂交技术在人类巨噬细胞cDNA文库中筛选到了一个新的COFILIN相互作用蛋白GOSR1(Golgi SNAP receptor complex member 1),然后采用GST pulldown和Co-IP技术验证了这两个蛋白在体外和体内相互作用的特异性,并通过将构建的截短突变体转化宿主菌表达的形式确定了相互作用的区段.将烟曲霉COFILIN基因在HEK293T细胞中过表达,结果发现COFILIN对细胞内源性GOSR1在转录水平和蛋白水平的表达都有一定的抑制作用,同时抑制细胞增殖、促进凋亡,并引起G2/M期阻滞.

Aspergillus fumigatus COFILIN is an actin depolymerizing factor in eukaryote,which can promote cytoskeletal dynamics.COFILIN protein not only participate in the process of infecting host cells,but also helped host cells escaping from immune system defenses.In order to explore the functions of COFILIN in host cells,researchers firstly found COFILIN could interact with GOSR1（Golgi SNAP receptor complex member 1）through yeast two-hybrid screening.Then by using GST pull-down and Co-IP assay,researchers confirmed the specific interaction between the two proteins in vitro and in vivo,and identified the interaction domain by constructing deletion mutants.By analyzing the expression of GOSR1 in HEK293 Tcells,researchers concluded that exogenous COFILIN could suppress the expression of GOSR1 in mRNA and protein level.Besides,it could inhibits cell proliferation,promote cell apoptosis,and causes G2/M block.