LcrV of Yersinia pestis enters infected eukaryotic cells by a virulence plasmid-independent mechanism

Kenneth A. Fields, Susan C. Straley

Research output: Contribution to journalArticle

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Abstract

Yersinia pestis is the causative agent of bubonic plague and possesses a set of plasmid-encoded, secretable virulence proteins termed LcrV and Yops which are essential for survival in mammalian hosts. Yops and LcrV are secreted by a type III mechanism (Ysc), and Yops are unidirectionally targeted into the cytosol of associated eukaryotic cells in a tissue culture infection model. LcrV is required for Yops targeting, and recent findings have revealed that it can localize to the bacterial surface; however, its fate in this infection model has not been investigated in detail. In this study, we compared the localization of LcrV to that of the targeted proteins YopE and YopM by immunoblot analysis of fractions of Yersinia-infected HeLa cultures or by laser-scanning confocal microscopy of infected monolayers. Both LcrV and YopE were secreted by contact-activated, extracellularly localized yersiniae and were targeted to the HeLa cell cytosol. Although a significant amount of LcrV partitioned to the culture medium (unlike YopE), this extracellular pool of LcrV was not the source of the LcrV that entered HeLa cells. Unlike targeting of YopE and YopM, targeting of LcrV occurred in the absence of a functional Ysc apparatus and other virulence plasmid (pCD1)- expressed proteins. However, the Ysc is necessary for LcrV to be released into the medium, and our recent work has shown that localization of LcrV on the bacterial surface requires the Ysc. These results indicate that two mechanisms exist for the secretion of LcrV by Y. pestis, both of which are activated by contact with eukaryotic cells. LcrV secreted by the Ysc reaches the bacterial surface and the surrounding medium, whereas the second is a novel, Ysc-independent pathway which results in localization of LcrV in the cytosol of infected cells but not the surrounding medium.

Original languageEnglish
Pages (from-to)4801-4813
Number of pages13
JournalInfection and Immunity
Volume67
Issue number9
StatePublished - Sep 1 1999

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Yersinia pestis
Eukaryotic Cells
Cytosol
Virulence
Yersinia
Plasmids
HeLa Cells
Proteins
Plague
Infection
Confocal Microscopy
Culture Media

ASJC Scopus subject areas

  • Immunology

Cite this

LcrV of Yersinia pestis enters infected eukaryotic cells by a virulence plasmid-independent mechanism. / Fields, Kenneth A.; Straley, Susan C.

In: Infection and Immunity, Vol. 67, No. 9, 01.09.1999, p. 4801-4813.

Research output: Contribution to journalArticle

Fields, Kenneth A. ; Straley, Susan C. / LcrV of Yersinia pestis enters infected eukaryotic cells by a virulence plasmid-independent mechanism. In: Infection and Immunity. 1999 ; Vol. 67, No. 9. pp. 4801-4813.
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