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Originally published In Press as doi:10.1074/jbc.M002467200 on August 4, 2000

J. Biol. Chem., Vol. 275, Issue 47, 36869-36875, November 24, 2000
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LcrV, a Substrate for Yersinia enterocolitica Type III Secretion, Is Required for Toxin Targeting into the Cytosol of HeLa Cells*

Vincent T. LeeDagger , Christina Tam, and Olaf Schneewind§

From the Department of Microbiology and Immunology, University of California Los Angeles School of Medicine, Los Angeles, California 90095

Pathogenic Yersinia species employ type III machines to transport virulence factors across the bacterial envelope. Some substrates for the type III machinery are secreted into the extracellular medium, whereas others are targeted into the cytosol of host cells. We found that during infection of tissue culture cells, yersiniae secrete small amounts of LcrV into the extracellular medium. Knockout mutations of lcrV abolish Yersinia targeting and reduce expression of the lcrGVHyopBD operon. In contrast, a block in LcrV secretion does not affect targeting, but results in premature expression and secretion of Yop proteins into the extracellular medium. LcrV-mediated activation of the type III pathway is thought to occur by sequestration of the regulatory factor LcrG, presumably via the formation of LcrV·LcrG complexes. These results suggest that intrabacterial LcrV regulates the expression and targeting of Yop proteins during Yersinia infection, whereas secreted LcrV is required to ensure specificity of Yop injection into eukaryotic cells.

* This work was supported in part by United States Public Health Service Grant AI 42797.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Supported by a fellowship from the National Science Foundation.

§ To whom correspondence should be addressed: Dept. of Microbiology and Immunology, UCLA School of Medicine, 10833 Le Conte Ave., Los Angeles, CA 90095. Tel.: 310-206-0997; Fax: 310-267-0173; E-mail: olafs@ucla.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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