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PMID: 15731026 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The StcE protease contributes to intimate adherence of enterohemorrhagic Escherichia coli O157:H7 to host cells.

Infection and immunity ·Vol. 73 ·No. 3 ·2005-03-00 ·Pages 1295-303

Grys TE, Siegel MB, Lathem WW, Welch RA

Abstract

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a diarrheal pathogen that causes attaching and effacing (A/E) lesions on intestinal epithelial cells. Strains of the O157 serogroup carry the large virulence plasmid pO157, which encodes the etp type II secretion system that secretes the genetically linked zinc metalloprotease StcE. The Ler regulator controls expression of many genes involved in A/E lesion formation, as well as StcE, suggesting StcE may be important at a similar time during colonization. Our laboratory has previously demonstrated that StcE cleaves C1-esterase inhibitor, a regulator of multiple inflammation pathways. Here we report two new substrates for StcE, mucin 7 and glycoprotein 340, and that purified StcE reduces the viscosity of human saliva. We tested the hypothesis that StcE contributes to intimate adherence of EHEC to host cells by cleavage of glycoproteins from the cell surface. The fluorescent actin stain (FAS) test was used to observe the intimate adherence represented by fluorescently stained bacteria colocalized with regions of bundled actin formed on HEp-2 cells. An E. coli O157:H7 strain with a stcE gene deletion was not affected in its ability to generally adhere to HEp-2 cells, but it did score threefold lower on the FAS test than wild-type or complemented strains. Addition of exogenous recombinant StcE increased intimate adherence of the mutant to wild-type levels. Thus, StcE may help block host clearance of E. coli O157:H7 by destruction of some classes of glycoproteins, and it contributes to intimate adherence of E. coli O157:H7 to the HEp-2 cell surface.

MeSH Terms
Bacterial Adhesion Cell Line Escherichia coli O157/enzymology,pathogenicity,physiology Escherichia coli Proteins/chemistry,genetics,metabolism Humans Metalloendopeptidases/chemistry,genetics,metabolism Molecular Sequence Data Mucins/metabolism Receptors, Immunologic/metabolism Saliva/physiology Salivary Proteins and Peptides Sequence Analysis, DNA Virulence
Chemicals
Escherichia coli Proteins MUC7 protein, human Mucins Receptors, Immunologic Salivary Proteins and Peptides glycoprotein 340 Metalloendopeptidases StcE protein, E coli
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Grys Thomas E
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, 1300 University Ave., Room 481 MSC, Madison, WI 53706, USA.
Siegel Matthew B
Lathem Wyndham W
Welch Rodney A
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.73.3.1295-1303.2005
Published
2005-03-00
Pages
1295-303
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC1064933
Subset
IM
Grants
NIAID NIH HHS · R01 AI051735 · United States
NIDDK NIH HHS · R01 DK063250 · United States
NIGMS NIH HHS · T32 GM008349 · United States
NIGMS NIH HHS · 5T32GM08349 · United States
Databases
GENBANK
AY074613, AY714880
External Links
PubMed source DOI: 10.1128/IAI.73.3.1295-1303.2005 PMC Full Text View journal details
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