کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3417313 1225200 2006 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
The Ser/Thr kinase activity of the Yersinia protein kinase A (YpkA) is necessary for full virulence in the mouse, mollifying phagocytes, and disrupting the eukaryotic cytoskeleton
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی میکروب شناسی
پیش نمایش صفحه اول مقاله
The Ser/Thr kinase activity of the Yersinia protein kinase A (YpkA) is necessary for full virulence in the mouse, mollifying phagocytes, and disrupting the eukaryotic cytoskeleton
چکیده انگلیسی

The Yersinia protein kinase A (YpkA) is injected into host cells by the yersinial type three secretion system (TTSS). YpkA is widely believed to function within the host cell based on the fact that its kinase domain is clearly homologous to eukaryotic Ser/Thr kinases and that its enzymatic activity, when assayed in vitro, is dependent on eukaryotic-derived host factors. Whether this activity is required for virulence has not been addressed. Here, we report that a Yersinia pseudotuberculosis strain expressing a kinase-inactive YpkAD270A variant is greatly attenuated in the mouse model of infection compared to the isogenic wild-type strain. The ypkAD270A mutant strain was likewise attenuated in a cell culture infection assay indicating that the kinase activity of YpkA enhances the viability of host cell-associated bacteria. To begin to understand what cellular activities are targeted, we expressed YpkA and its variants in two different yeast model systems. In agreement with previous studies, we found that when rapidly induced and expressed at high levels in Saccharomyces cerevisiae, YpkA-mediated toxicity occurred extremely swiftly. Under these conditions toxicity was dependent on the structurally distinct GTPase-binding domain of YpkA and was entirely independent of its kinase activity. Therefore, to probe for kinase-dependent effects we expressed YpkA and its kinase-inactive variant at comparatively moderate levels in the fission yeast Schizosaccharomyces pombe. S. pombe is particularly well suited for actin cytoskeletal studies due to its easily quantifiable, well defined pattern of actin localization. S. pombe transformed with a wild-type YpkA-encoding plasmid displayed a pronounced actin mislocalization phenotype, the severity of which was directly proportional to the level of YpkA expressed in the cell. In cells expressing the kinase-inactive YpkA variant, the degree of actin mislocalization was reduced, but not entirely abrogated, suggesting that YpkA affects the eukaryotic cytoskeleton through kinase-dependent and kinase-independent mechanisms. Collectively, our yeast-derived results show how critical expression levels and exposure periods are for assaying virulence factor activities in heterologous model systems. More generally, our finding that the ‘eukaryotic-like’ kinase domain of YpkA is important for virulence illustrates how a bacterium can utilize a host-like factor or activity in order to enhance its survival following host cell contact.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Microbial Pathogenesis - Volume 40, Issue 5, May 2006, Pages 234–243
نویسندگان
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