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Infection and Immunity, March 1999, p. 1359-1367, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Expression of the Plague Plasminogen Activator in Yersinia pseudotuberculosis and Escherichia coli

V. Kutyrev,1 R. J. Mehigh,2,dagger V. L. Motin,2,Dagger M. S. Pokrovskaya,3 G. B. Smirnov,3 and R. R. Brubaker2,*

Laboratory of Molecular Microbiology, Russian Research Anti-Plague Institute "Microbe," Saratov 410071,1 and Gamaleya Research Institute for Epidemiology and Microbiology, Moscow 123098,3 Russia, and Department of Microbiology, Michigan State University, East Lansing, Michigan 488242

Received 6 August 1998/Returned for modification 28 September 1998/Accepted 7 December 1998

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 ~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.

* Corresponding author. Mailing address: Department of Microbiology, 57A Giltner Hall, Michigan State University, East Lansing, MI 48824-1101. Phone: (517) 355-6466. Fax: (517) 353-8957. E-mail: brubake3{at}pilot.msu.edu.

dagger Present address: Sigma Chemical Company, St. Louis, MO 63178.

Dagger Present address: Lawrence Livermore Lab, Livermore, CA 94550.

Infection and Immunity, March 1999, p. 1359-1367, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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