Kutyrev V, Mehigh R J, Motin V L, Pokrovskaya M S, Smirnov G B, Brubaker R R
Laboratory of Molecular Microbiology, Russian Research Anti-Plague Institute "Microbe," Saratov 410071, Russia.
Infect Immun. 1999 Mar;67(3):1359-67. doi: 10.1128/IAI.67.3.1359-1367.1999.
Enteropathogenic yersiniae (Yersinia pseudotuberculosis and Yersinia enterocolitica) typically cause chronic disease as opposed to the closely related Yersinia pestis, the causative agent of bubonic plague. It is established that this difference reflects, in part, carriage by Y. pestis of a unique 9.6-kb pesticin or Pst plasmid (pPCP) encoding plasminogen activator (Pla) rather than distinctions between shared approximately 70-kb low-calcium-response, or Lcr, plasmids (pCD in Y. pestis and pYV in enteropathogenic yersiniae) encoding cytotoxic Yops and anti-inflammatory V antigen. Pla is known to exist as a combination of 32.6-kDa (alpha-Pla) and slightly smaller (beta-Pla) outer membrane proteins, of which at least one promotes bacterial dissemination in vivo and degradation of Yops in vitro. We show here that only alpha-Pla accumulates in Escherichia coli LE392/pPCP1 cultivated in enriched medium and that either autolysis or extraction of this isolate with 1.0 M NaCl results in release of soluble alpha and beta forms possessing biological activity. This process also converted cell-bound alpha-Pla to beta-Pla and smaller forms in Y. pestis KIM/pPCP1 and Y. pseudotuberculosis PB1/+/pPCP1 but did not promote solubilization. Pla-mediated posttranslational hydrolysis of pulse-labeled Yops in Y. pseudotuberculosis PB1/+/pPCP1 occurred more slowly than that in Y. pestis but was otherwise similar except for accumulation of stable degradation products of YadA, a pYV-mediated fibrillar adhesin not encoded in frame by pCD. Carriage of pPCP by Y. pseudotuberculosis did not significantly influence virulence in mice.
肠道致病性耶尔森菌(假结核耶尔森菌和小肠结肠炎耶尔森菌)通常引起慢性病,这与密切相关的鼠疫耶尔森菌不同,鼠疫耶尔森菌是腺鼠疫的病原体。已经确定,这种差异部分反映了鼠疫耶尔森菌携带独特的9.6kb杀鼠疫菌素或Pst质粒(pPCP),该质粒编码纤溶酶原激活剂(Pla),而不是共享的约70kb低钙反应或Lcr质粒(鼠疫耶尔森菌中的pCD和肠道致病性耶尔森菌中的pYV)之间的区别,这些质粒编码细胞毒性Yops和抗炎V抗原。已知Pla以32.6kDa(α-Pla)和稍小(β-Pla)的外膜蛋白组合形式存在,其中至少一种促进细菌在体内的传播和体外Yops的降解。我们在此表明,在富集培养基中培养的大肠杆菌LE392/pPCP1中仅积累α-Pla,并且该分离株的自溶或用1.0M NaCl提取导致具有生物活性的可溶性α和β形式的释放。该过程还将鼠疫耶尔森菌KIM/pPCP1和假结核耶尔森菌PB1/+/pPCP1中细胞结合的α-Pla转化为β-Pla和更小的形式,但不促进溶解。假结核耶尔森菌PB1/+/pPCP1中Pla介导的脉冲标记Yops的翻译后水解比鼠疫耶尔森菌中发生得更慢,但除此之外相似,除了YadA的稳定降解产物的积累,YadA是一种由pYV介导的纤维状粘附素,未由pCD框内编码。假结核耶尔森菌携带pPCP对小鼠的毒力没有显著影响。