Wright A C, Powell J L, Kaper J B, Morris J G
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
Infect Immun. 2001 Nov;69(11):6893-901. doi: 10.1128/IAI.69.11.6893-6901.2001.
Virulence of Vibrio vulnificus correlates with changes in colony morphology that are indicative of a reversible phase variation for expression of capsular polysaccharide (CPS). Encapsulated variants are virulent with opaque colonies, whereas phase variants with reduced CPS expression are attenuated and are translucent. Using TnphoA mutagenesis, we identified a V. vulnificus CPS locus, which included an upstream ops element, a wza gene (wza(Vv)), and several open reading frames with homology to CPS biosynthetic genes. This genetic organization is characteristic of group 1 CPS operons. The wza gene product is required for transport of CPS to the cell surface in Escherichia coli. Polar transposon mutations in wza(Vv) eliminated expression of downstream biosynthetic genes, confirming operon structure. On the other hand, nonpolar inactivation of wza(Vv) was specific for CPS transport, did not alter CPS biosynthesis, and could be complemented in trans. Southern analysis of CPS phase variants revealed deletions or rearrangements at this locus. A survey of environmental isolates indicated a correlation between deletions in wza(Vv) and loss of virulent phenotype, suggesting a genetic mechanism for CPS phase variation. Full virulence in mice required surface expression of CPS and supported the essential role of capsule in the pathogenesis of V. vulnificus.
创伤弧菌的毒力与菌落形态的变化相关,这些变化表明其荚膜多糖(CPS)表达存在可逆的相变。被包裹的变体具有不透明菌落且具有毒力,而CPS表达降低的相变变体则毒力减弱且呈半透明状。通过TnphoA诱变,我们鉴定出了一个创伤弧菌CPS基因座,其中包括一个上游操纵子元件、一个wza基因(wza(Vv))以及几个与CPS生物合成基因具有同源性的开放阅读框。这种基因组织是1型CPS操纵子的特征。wza基因产物是大肠杆菌中CPS转运至细胞表面所必需的。wza(Vv)中的极性转座子突变消除了下游生物合成基因的表达,证实了操纵子结构。另一方面,wza(Vv)的非极性失活对CPS转运具有特异性,不会改变CPS生物合成,并且可以通过反式互补。对CPS相变变体的Southern分析揭示了该基因座处的缺失或重排。对环境分离株的调查表明,wza(Vv)中的缺失与毒力表型的丧失之间存在相关性,提示了CPS相变的遗传机制。小鼠体内的完全毒力需要CPS的表面表达,并支持了荚膜在创伤弧菌致病过程中的重要作用。