Monack D M, Mecsas J, Ghori N, Falkow S
Department of Microbiology and Immunology, Stanford School of Medicine, Stanford University, Stanford CA 94305, USA.
Proc Natl Acad Sci U S A. 1997 Sep 16;94(19):10385-90. doi: 10.1073/pnas.94.19.10385.
Pathogenic Yersinia spp. carry a large common plasmid that encodes a number of essential virulence determinants. Included in these factors are the Yersinia-secreted proteins called Yops. We analyzed the consequences of wild-type and mutant strains of Yersinia pseudotuberculosis interactions with the macrophage cell line RAW264. 7 and murine bone marrow-derived macrophages. Wild-type Y. pseudotuberculosis kills approximately 70% of infected RAW264.7 macrophages and marrow-derived macrophages after an 8-h infection. We show that the cell death mediated by Y. pseudotuberculosis is apoptosis. Mutant Y. pseudotuberculosis that do not make any Yop proteins no longer cause host cell death. Attachment to host cells via invasin or YadA is necessary for the cell death phenotype. Several Yop mutant strains that fail to express one or more Yop proteins were engineered and then characterized for their ability to cause host cell death. A mutant with a polar insertion in YpkA Ser/Thr kinase that does not express YpkA or YopJ is no longer able to cause apoptosis. In contrast, a mutant no longer making YopE or YopH (a tyrosine phosphatase) induces apoptosis in macrophages similar to wild type. When yopJ is added in trans to the ypkAyopJ mutant, the ability of this strain to signal programmed cell death in macrophages is restored. Thus, YopJ is necessary for inducing apoptosis. The ability of Y. pseudotuberculosis to promote apoptosis of macrophages in cell culture suggests that this process is important for the establishment of infection in the host and for evasion of the host immune response.
致病性耶尔森菌携带一个大型共质体,该共质体编码许多必需的毒力决定因素。这些因素包括被称为Yops的耶尔森菌分泌蛋白。我们分析了野生型和突变型假结核耶尔森菌菌株与巨噬细胞系RAW264.7和小鼠骨髓来源巨噬细胞相互作用的后果。野生型假结核耶尔森菌在感染8小时后可杀死约70%被感染的RAW264.7巨噬细胞和骨髓来源巨噬细胞。我们表明,假结核耶尔森菌介导的细胞死亡是凋亡。不产生任何Yop蛋白的突变型假结核耶尔森菌不再导致宿主细胞死亡。通过侵袭素或YadA附着于宿主细胞是细胞死亡表型所必需的。构建了几种未能表达一种或多种Yop蛋白的Yop突变株,然后对其导致宿主细胞死亡的能力进行了表征。YpkA丝氨酸/苏氨酸激酶中存在极性插入且不表达YpkA或YopJ的突变体不再能够诱导凋亡。相比之下,不再产生YopE或YopH(一种酪氨酸磷酸酶)的突变体在巨噬细胞中诱导凋亡的情况与野生型相似。当将yopJ反式添加到ypkAyopJ突变体中时,该菌株在巨噬细胞中引发程序性细胞死亡的信号传导能力得以恢复。因此,YopJ是诱导凋亡所必需的。假结核耶尔森菌在细胞培养中促进巨噬细胞凋亡的能力表明,这一过程对于在宿主体内建立感染以及逃避宿主免疫反应很重要。