During Russell L, Gibson Bruce G, Li Wei, Bishai Ellen A, Sidhu Gurjit S, Landry Jacques, Southwick Frederick S
Department of Medicine, University of Florida College of Medicine, Gainesville, FL 32610, USA.
EMBO J. 2007 May 2;26(9):2240-50. doi: 10.1038/sj.emboj.7601687. Epub 2007 Apr 19.
Inhalation of anthrax causes fatal bacteremia, indicating a meager host immune response. We previously showed that anthrax lethal toxin (LT) paralyzes neutrophils, a major component of innate immunity. Here, we have found that LT also inhibits actin-based motility of the intracellular pathogen Listeria monocytogenes. LT inhibition of actin assembly is mediated by blockade of Hsp27 phosphorylation, and can be reproduced by treating cells with the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580. Nonphosphorylated Hsp27 inhibits Listeria actin-based motility in cell extracts, and binds to and sequesters purified actin monomers. Phosphorylation of Hsp27 reverses these effects. RNA interference knockdown of Hsp27 blocks LT inhibition of Listeria actin-based motility. Rescue with wild-type Hsp27 accelerates Listeria speed in knockdown cells, whereas introduction of Hsp27 mutants incapable of phosphorylation or dephosphorylation causes slowing down. We propose that Hsp27 facilitates actin-based motility through a phosphorylation cycle that shuttles actin monomers to regions of new actin filament assembly. Our findings provide a previously unappreciated mechanism for LT virulence, and emphasize a central role for p38 MAP kinase-mediated phosphorylation of Hsp27 in actin-based motility and innate immunity.
吸入炭疽会导致致命的菌血症,这表明宿主的免疫反应微弱。我们之前发现炭疽致死毒素(LT)会使中性粒细胞麻痹,中性粒细胞是固有免疫的主要组成部分。在此,我们发现LT还会抑制细胞内病原体单核细胞增生李斯特菌基于肌动蛋白的运动。LT对肌动蛋白组装的抑制作用是通过阻断Hsp27磷酸化介导的,用p38丝裂原活化蛋白(MAP)激酶抑制剂SB203580处理细胞也能产生同样的效果。未磷酸化的Hsp27在细胞提取物中会抑制李斯特菌基于肌动蛋白的运动,并与纯化的肌动蛋白单体结合并使其隔离。Hsp27的磷酸化会逆转这些效应。通过RNA干扰敲低Hsp27会阻断LT对李斯特菌基于肌动蛋白运动的抑制作用。用野生型Hsp27进行拯救会加快敲低细胞中李斯特菌的运动速度,而引入无法磷酸化或去磷酸化的Hsp27突变体则会导致其运动速度减慢。我们提出Hsp27通过一个磷酸化循环促进基于肌动蛋白的运动,该循环将肌动蛋白单体转运到新肌动蛋白丝组装的区域。我们的研究结果揭示了一种此前未被认识的LT毒力机制,并强调了p38 MAP激酶介导的Hsp27磷酸化在基于肌动蛋白的运动和固有免疫中的核心作用。