Tang P, Sutherland C L, Gold M R, Finlay B B
Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.
Infect Immun. 1998 Mar;66(3):1106-12. doi: 10.1128/IAI.66.3.1106-1112.1998.
PD98059, a specific inhibitor of MEK-1 mitogen-activated protein (MAP) kinase kinase, blocked Listeria monocytogenes invasion into HeLa epithelial cells. The effects of PD98059 were reversible, as adherent extracellular bacteria were internalized upon removal of the drug. Previously, we reported that L. monocytogenes could activate ERK-1 and ERK-2 MAP kinases through the action of listeriolysin O (LLO) on the host cell (P. Tang, I. Rosenshine, P. Cossart, and B. B. Finlay, Infect. Immun. 64:2359-2361, 1996). We have now found that two other MAP kinase pathways, those of p38 MAP kinase and c-Jun N-terminal kinase, are also activated by wild-type L. monocytogenes. Mutants lacking functional LLO (hly mutants) were still invasive but only activated ERK-2 and only activated it at later (90-min) postinfection times. Two inhibitors of L. monocytogenes invasion, cytochalasin D, which disrupts actin polymerization, and wortmannin, which blocks phosphatidylinositol (PI) 3-kinase activity, did not block ERK-2 activation by wild-type L. monocytogenes and hly mutants. However, genistein, an inhibitor of tyrosine kinases, and PD98059 both blocked invasion and decreased ERK-2 activation. These results suggest that MEK-1 and ERK-2 activities are essential for L. monocytogenes invasion into host epithelial cells. This is the first report to show that a MAP kinase pathway is required for bacterial invasion.
PD98059是丝裂原活化蛋白(MAP)激酶激酶MEK-1的特异性抑制剂,它能阻断单核细胞增生李斯特菌侵入HeLa上皮细胞。PD98059的作用是可逆的,因为去除药物后,附着在细胞外的细菌会被内化。此前,我们报道过单核细胞增生李斯特菌可通过溶血素O(LLO)对宿主细胞的作用激活ERK-1和ERK-2 MAP激酶(P. Tang、I. Rosenshine、P. Cossart和B. B. Finlay,《感染与免疫》64:2359 - 2361,1996年)。我们现在发现,另外两条MAP激酶途径,即p38 MAP激酶和c - Jun N端激酶途径,也会被野生型单核细胞增生李斯特菌激活。缺乏功能性LLO的突变体(hly突变体)仍具有侵袭性,但仅激活ERK-2,且仅在感染后较晚时间(90分钟)激活。两种单核细胞增生李斯特菌侵袭抑制剂,即破坏肌动蛋白聚合的细胞松弛素D和阻断磷脂酰肌醇(PI)3激酶活性的渥曼青霉素,均不能阻断野生型单核细胞增生李斯特菌和hly突变体对ERK-2的激活。然而,酪氨酸激酶抑制剂染料木黄酮和PD98059都能阻断侵袭并降低ERK-2的激活。这些结果表明,MEK-1和ERK-2的活性对于单核细胞增生李斯特菌侵入宿主上皮细胞至关重要。这是首次报道表明MAP激酶途径是细菌侵袭所必需的。