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Infection and Immunity, January 1999, p. 120-125, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Expression of the EspB Protein of Enteropathogenic
Escherichia coli within HeLa Cells Affects Stress Fibers and
Cellular Morphology
Kathleen A.
Taylor,1
Paul W.
Luther,2 and
Michael S.
Donnenberg1,*
Division of Infectious Diseases, Department
of Medicine,1 and
Department of
Physiology,2 University of Maryland School of
Medicine, Baltimore, Maryland 21201
Received 13 July 1998/Returned for modification 6 October
1998/Accepted 28 October 1998
The EspB protein of enteropathogenic Escherichia coli
(EPEC) is essential for the signaling events that lead to the
accumulation of actin beneath intimately attached bacteria, a process
that is known as the attaching and effacing effect. EspB is targeted to
the host cell cytoplasm by a type III secretion apparatus. To determine
the effect of intracellular EspB on the host cell cytoskeleton, we
transfected HeLa cells with a plasmid containing the espB
gene under the control of an inducible eukaryotic promoter. A HeLa cell
clone that expressed espB mRNA and EspB protein after induction was selected for further study. The expression of EspB in
these cells caused a dramatic change in cell morphology and a marked
reduction in actin stress fibers. Cells expressing EspB were
significantly impaired in their ability to support invasion by EPEC and
Salmonella typhimurium. However, the expression of EspB
within host cells could not compensate for the lack of EspB expression
by an espB mutant strain of EPEC to restore attaching and
effacing activity. These studies suggest that EspB is a cytoskeletal toxin that is translocated to the host cell cytoplasm, where it causes
a redistribution of actin.
*
Corresponding author. Mailing address: Division of
Infectious Diseases, University of Maryland School of Medicine, 10 South Pine St., MSTF 900, Baltimore, MD 21201. Phone: (410) 706-7560. Fax: (410) 706-8700. E-mail: mdonnenb{at}umaryland.edu.
Infection and Immunity, January 1999, p. 120-125, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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