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鼠疫耶尔森菌外部蛋白E(YopE)是鼠疫耶尔森菌的6种分泌蛋白之一,主要通过其144位的“精氨酸手指”结构与细胞膜耦联蛋白RhoGTP酶相互作用发挥功能.本文构建YopE及其144位突变体YopE(R144A)的可诱导表达系统,并优化了诱导条件.用该系统结合流式细胞技术检测YopE和YopE(R144A)对细胞凋亡、细胞周期和细胞活性氧(ROS)水平的影响.结果显示:YopE(R144A)促进HeLa细胞凋亡;使G0/G1期细胞比例上升,G2/M期细胞比例下降;随着YopE(R144A)表达量增加,p21蛋白的表达量也增加;YopE(R144A)也能抑制细胞ROS的产生.研究结果提示,YopE在细胞内可能存在新的致病靶点.
Yersinia pestis YE is one of the six secretory proteins of Yersinia pestis and plays its role mainly through the interaction of its 144 “arginine finger” structure with the membrane-associated protein RhoGTPase Function.In this paper, an inducible expression system of YopE and its 144 mutant YopE (R144A) was constructed and the conditions of induction were optimized.Using this system combined with flow cytometry to detect the effects of YopE and YopE (R144A) on apoptosis, cell cycle and The results showed that YopE (R144A) promoted the apoptosis of HeLa cells, increased the proportion of cells in G0 / G1 phase and decreased the proportion of cells in G2 / M phase. With the increase of YopE (R144A) The expression of p21 protein also increased.YopE (R144A) also inhibited the production of ROS.The results suggest that YopE may have new target in the cell.