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首页> 外文期刊>BMC Microbiology >Yersinia outer protein YopE affects the actin cytoskeleton in Dictyostelium discoideum through targeting of multiple Rho family GTPases
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Yersinia outer protein YopE affects the actin cytoskeleton in Dictyostelium discoideum through targeting of multiple Rho family GTPases

机译：耶尔森氏菌外蛋白YopE通过靶向多个Rho家族的GTPases影响Disctyostelium Discoideum中的肌动蛋白细胞骨架

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Background All human pathogenic Yersinia species share a virulence-associated type III secretion system that translocates Yersinia effector proteins into host cells to counteract infection-induced signaling responses and prevent phagocytosis. Dictyostelium discoideum has been recently used to study the effects of bacterial virulence factors produced by internalized pathogens. In this study we explored the potential of Dictyostelium as model organism for analyzing the effects of ectopically expressed Yersinia outer proteins (Yops). Results The Yersinia pseudotuberculosis virulence factors YopE, YopH, YopM and YopJ were expressed de novo within Dictyostelium and their effects on growth in axenic medium and on bacterial lawns were analyzed. No severe effect was observed for YopH, YopJ and YopM, but expression of YopE, which is a GTPase activating protein for Rho GTPases, was found to be highly detrimental. GFP-tagged YopE expressing cells had less conspicuous cortical actin accumulation and decreased amounts of F-actin. The actin polymerization response upon cAMP stimulation was impaired, although chemotaxis was unaffected. YopE also caused reduced uptake of yeast particles. These alterations are probably due to impaired Rac1 activation. We also found that YopE predominantly associates with intracellular membranes including the Golgi apparatus and inhibits the function of moderately overexpressed RacH. Conclusion The phenotype elicited by YopE in Dictyostelium can be explained, at least in part, by inactivation of one or more Rho family GTPases. It further demonstrates that the social amoeba Dictyostelium discoideum can be used as an efficient and easy-to-handle model organism in order to analyze the function of a translocated GAP protein of a human pathogen.
机译：背景技术所有人类致病性耶尔森氏菌物种均具有与毒性相关的III型分泌系统，该系统将耶尔森氏菌效应蛋白转运到宿主细胞中，以抵消感染诱导的信号传导反应并防止吞噬作用。盘基网柄菌最近已用于研究由内在病原体产生的细菌毒力因子的作用。在这项研究中，我们探索了Dictyostelium作为模型生物体的潜力，可用于分析异位表达的耶尔森氏菌外蛋白（Yops）的作用。结果鼠疫耶尔森菌假结核耶尔森氏菌毒力因子YopE，YopH，YopM和YopJ从头到尾表达，并分析了它们在无菌培养基和细菌性草坪中的生长。没有观察到对YopH，YopJ和YopM的严重影响，但是发现作为Rho GTPases的GTP酶激活蛋白的YopE的表达是高度有害的。 GFP标记的YopE表达细胞的皮层肌动蛋白积累不那么明显，F-肌动蛋白的数量减少。尽管趋化性不受影响，但cAMP刺激后的肌动蛋白聚合反应受到损害。 YopE还引起酵母颗粒摄取减少。这些改变可能是由于Rac1激活受损。我们还发现，YopE主要与包括高尔基体在内的细胞内膜缔合，并抑制中度过量表达的RacH的功能。结论YopE在双歧杆菌中产生的表型可以至少部分地通过一种或多种Rho家族GTPases的失活来解释。它进一步证明社交变形虫盘基网柄线虫可以用作有效且易于处理的模型生物，以便分析人类病原体的易位GAP蛋白的功能。

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