Swanson Kena A, Crane Deborah D, Caldwell Harlan D
Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 903 S. 4th St., Hamilton, MT 59840, USA.
Infect Immun. 2007 Dec;75(12):5669-77. doi: 10.1128/IAI.01096-07. Epub 2007 Oct 1.
Chlamydia trachomatis is an obligate intracellular pathogen of humans that exhibits species-specific biological characteristics in its early interactions with host cells that are likely important to pathogenesis. One such characteristic is the tyrosine phosphorylation (Tyr-P) of an approximately 70-kDa polypeptide that occurs only after infection of mammalian cells by human strains. We sought to identify this protein because of its potential significance to the pathogenesis of human chlamydial infections. Using an immunoproteomic approach we identified the host protein ezrin, a member of the ezrin-radixin-moesin (ERM) protein family that serves as a physical link between host cell receptors and the actin cytoskeleton. Confocal microscopy studies showed colocalization of ezrin and actin at the tips and crypts of microvilli, the site of chlamydial attachment and entry, respectively. To demonstrate a functional role for ezrin we infected cells with a dominant-negative (DN) ezrin phenotype or treated cells with ezrin-specific small interfering RNA (siRNA). We found that both DN and siRNA-treated cells were significantly less susceptible to infection by human chlamydial strains. Moreover, we demonstrated that inhibition of infection in ezrin DN cells occurred at the stage of chlamydial entry. We hypothesize that the C. trachomatis-specific Tyr-P of ezrin might relate to an undefined species-specific mechanism of pathogen entry that involves chlamydial specific ligand(s) and host cell coreceptor usage.
沙眼衣原体是人类专性胞内病原体,在其与宿主细胞的早期相互作用中表现出物种特异性生物学特性,这可能对发病机制很重要。其中一个特性是一种约70 kDa多肽的酪氨酸磷酸化(Tyr-P),这种磷酸化仅在人源菌株感染哺乳动物细胞后才会发生。由于其对人类衣原体感染发病机制的潜在重要性,我们试图鉴定这种蛋白质。我们采用免疫蛋白质组学方法鉴定出宿主蛋白埃兹蛋白,它是埃兹蛋白-根蛋白-膜突蛋白(ERM)家族的成员,在宿主细胞受体和肌动蛋白细胞骨架之间起物理连接作用。共聚焦显微镜研究显示,埃兹蛋白和肌动蛋白分别在微绒毛的尖端和隐窝处共定位,微绒毛分别是衣原体附着和进入的部位。为了证明埃兹蛋白的功能作用,我们用显性负性(DN)埃兹蛋白表型感染细胞,或用埃兹蛋白特异性小干扰RNA(siRNA)处理细胞。我们发现,DN细胞和经siRNA处理的细胞对人源衣原体菌株感染的敏感性均显著降低。此外,我们证明,埃兹蛋白DN细胞中感染的抑制发生在衣原体进入阶段。我们推测,沙眼衣原体特异性的埃兹蛋白Tyr-P可能与一种未明确的病原体进入的物种特异性机制有关,该机制涉及衣原体特异性配体和宿主细胞共受体的使用。