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Infection and Immunity, November 1999, p. 6019-6025, Vol. 67, No. 11
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Citrobacter rodentium espB Is Necessary for Signal Transduction and for Infection of Laboratory Mice

Joseph V. Newman,1 Brian A. Zabel,1,dagger Sharda S. Jha,1,Dagger and David B. Schauer1,2,*

Division of Bioengineering and Environmental Health1 and Division of Comparative Medicine,2 Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Received 10 May 1999/Returned for modification 23 July 1999/Accepted 20 August 1999

Citrobacter rodentium is the causative agent of transmissible murine colonic hyperplasia and contains a locus of enterocyte effacement (LEE) similar to that found in enteropathogenic Escherichia coli (EPEC). EPEC espB is necessary for intimate attachment and signal transduction between EPEC and cultured cell monolayers. Mice challenged with wild-type C. rodentium develop a mucosal immunoglobulin A response to EspB. In this study, C. rodentium espB has been cloned and its nucleotide sequence has been determined. C. rodentium espB was found to have 90% identity to EPEC espB. A nonpolar insertion mutation in C. rodentium espB was constructed and used to replace the chromosomal wild-type allele. The C. rodentium espB mutant exhibited reduced cell association and had no detectable fluorescent actin staining activity on cultured cell monolayers. The C. rodentium espB mutant also failed to colonize laboratory mice following experimental inoculation. The espB mutation could be complemented with a plasmid-encoded copy of the gene, which restored both cell association and fluorescent actin staining activity, as well as the ability to colonize laboratory mice. These studies indicate that espB is necessary for signal transduction and for colonization of laboratory mice by C. rodentium.

* Corresponding author. Mailing address: Room 56-787B, MIT, Cambridge, MA 02139. Phone: (617) 253-8113. Fax: (617) 258-0225. E-mail: schauer{at}mit.edu.

dagger Present address: LeukoSite, Inc., Cambridge, MA 02142.

Dagger Present address: Genzyme Corp., Framingham, MA 01710.

Infection and Immunity, November 1999, p. 6019-6025, Vol. 67, No. 11
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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