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Infection and Immunity, May 1999, p. 2389-2398, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Enterohemorrhagic Escherichia coli
O157:H7 Produces Tir, Which Is Translocated to the Host Cell
Membrane but Is Not Tyrosine Phosphorylated
Rebekah
DeVinney,
Markus
Stein,
Dieter
Reinscheid,
Akio
Abe,
Sharon
Ruschkowski, and
B. Brett
Finlay*
Biotechnology Laboratory, University of
British Columbia, Vancouver, British Columbia V6T 1Z3, Canada
Received 2 December 1998/Returned for modification 26 January
1999/Accepted 16 February 1999
Intimate attachment to the host cell leading to the formation of
attaching and effacing (A/E) lesions is an essential feature of
enterohemorrhagic Escherichia coli (EHEC) O157:H7
pathogenesis. In a related pathogen, enteropathogenic E. coli (EPEC), this activity is dependent upon translocation of the
intimin receptor, Tir, which becomes tyrosine phosphorylated within the
host cell membrane. In contrast, the accumulation of
tyrosine-phosphorylated proteins beneath adherent EHEC bacteria does
not occur, leading to questions about whether EHEC uses a Tir-based
mechanism for adherence and A/E lesion formation. In this report, we
demonstrate that EHEC produces a functional Tir that is inserted into
host cell membranes, where it serves as an intimin receptor. However,
unlike in EPEC, in EHEC Tir is not tyrosine phosphorylated yet plays a
key role in both bacterial adherence to epithelial cells and pedestal
formation. EHEC, but not EPEC, was unable to synthesize Tir in
Luria-Bertani medium but was able to secrete Tir into M9 medium,
suggesting that Tir synthesis and secretion may be regulated
differently in these two pathogens. EHEC Tir and EPEC Tir both bind
intimin and focus cytoskeletal rearrangements, indicating that tyrosine phosphorylation is not needed for pedestal formation. EHEC and EPEC
intimins are functionally interchangeable, but EHEC Tir shows a much
greater affinity for EHEC intimin than for EPEC intimin. These findings
highlight some of the differences and similarities between EHEC and
EPEC virulence mechanisms, which can be exploited to further define the
molecular basis of pedestal formation.
*
Corresponding author. Mailing address: Biotechnology
Laboratory, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada. Phone: (604) 822-2210. Fax: (604) 822-9830. E-mail: bfinlay{at}unixg.ubc.ca.

Present address: Via Fiorentina 11, 53000 Siena,
Italy.

Present address: Abteilung Angewandte Mikrobiologie und Mykologie,
Universitaet Ulm, D-89069 Ulm,
Germany.
Infection and Immunity, May 1999, p. 2389-2398, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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