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Originally published In Press as doi:10.1074/jbc.M112224200 on February 27, 2002

J. Biol. Chem., Vol. 277, Issue 18, 15607-15612, May 3, 2002
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Cytotoxic Necrotizing Factor-1 Contributes to Escherichia coli K1 Invasion of the Central Nervous System*

Naveed Ahmed KhanDagger , Ying Wang§, Kee Jun KimDagger , Jin Woong ChungDagger , Carol Ann Wass§, and Kwang Sik KimDagger §

From the § Division of Infectious Diseases, Childrens Hospital Los Angeles, University of Southern California School of Medicine, Los Angeles, California 90027 and the Dagger  Division of Pediatric Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287

Escherichia coli K1 invasion of brain microvascular endothelial cells (BMECs) is a prerequisite for penetration into the central nervous system and requires actin cytoskeletal rearrangements. Here, we demonstrate that E. coli K1 invasion of BMECs requires RhoA activation. In addition, we show that cytotoxic necrotizing factor-1 (CNF1) contributes to E. coli K1 invasion of brain endothelial cells in vitro and traversal of the blood-brain barrier in the experimental hematogenous meningitis animal model. These in vitro and in vivo effects of CNF1 were dependent upon RhoA activation as shown by (a) decreased invasion and RhoA activation with the Delta cnf1 mutant of E. coli K1 and (b) restoration of invasion frequency of the Delta cnf1 mutant to the level of the parent E. coli K1 strain in BMECs with constitutively active RhoA. In addition, CNF1-enhanced E. coli invasion of brain endothelial cells and stress fiber formation were independent of focal adhesion kinase and phosphatidylinositol 3-kinase activation. This is the first demonstration that CNF1 contributes to E. coli K1 invasion of BMECs.

* This work was supported by National Institutes of Health Grants NS 26310, AI 47225, and HL 61951.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.

To whom correspondence should be addressed: Division of Pediatric Infectious Diseases, Johns Hopkins University School of Medicine, 600 N. Wolfe St., Park 256, Baltimore, MD 21287. Tel.: 410-614-3917; Fax: 410-614-1491; E-mail: kwangkim@jhmi.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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