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PMID: 11119535 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Characterization of Vibrio cholerae O1 El tor galU and galE mutants: influence on lipopolysaccharide structure, colonization, and biofilm formation.

Infection and immunity ·Vol. 69 ·No. 1 ·2001-01-00 ·Pages 435-45

Nesper J, Lauriano CM, Klose KE, Kapfhammer D, Kraiss A, Reidl J

Abstract

Recently we described the isolation of spontaneous bacteriophage K139-resistant Vibrio cholerae O1 El Tor mutants. In this study, we identified phage-resistant isolates with intact O antigen but altered core oligosaccharide which were also affected in galactose catabolism; this strains have mutations in the galU gene. We inactivated another gal gene, galE, and the mutant was also found to be defective in the catabolism of exogenous galactose but synthesized an apparently normal lipopolysaccharide (LPS). Both gal mutants as well as a rough LPS (R-LPS) mutant were investigated for the ability to colonize the mouse small intestine. The galU and R-LPS mutants, but not the galE mutant, were defective in colonization, a phenotype also associated with O-antigen-negative mutants. By investigating several parameters in vitro, we could show that galU and R-LPS mutants were more sensitive to short-chain organic acids, cationic antimicrobial peptides, the complement system, and bile salts as well as other hydrophobic agents, indicating that their outer membrane no longer provides an effective barrier function. O-antigen-negative strains were found to be sensitive to complement and cationic peptides, but they displayed significant resistance to bile salts and short-chain organic acids. Furthermore, we found that galU and galE are essential for the formation of a biofilm in a spontaneous phage-resistant rugose variant, suggesting that the synthesis of UDP-galactose via UDP-glucose is necessary for biosynthesis of the exopolysaccharide. In addition, we provide evidence that the production of exopolysaccharide limits the access of phage K139 to its receptor, the O antigen. In conclusion, our results indicate involvement of galU in V. cholerae virulence, correlated with the observed change in LPS structure, and a role for galU and galE in environmental survival of V. cholerae.

MeSH Terms
Animals Bacterial Proteins/genetics,physiology Bile/physiology Biofilms Escherichia coli Proteins Fimbriae, Bacterial/physiology Galactose/metabolism Lipopolysaccharides/chemistry Mice Mutation O Antigens/physiology Open Reading Frames UDPglucose 4-Epimerase/genetics,physiology UTP-Glucose-1-Phosphate Uridylyltransferase Vibrio cholerae/genetics,immunology,physiology Virulence
Chemicals
Bacterial Proteins Escherichia coli Proteins Lipopolysaccharides O Antigens UTP-Glucose-1-Phosphate Uridylyltransferase galU protein, E coli UDPglucose 4-Epimerase Galactose
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nesper J
Zentrum für Infektionsforschung, Universität Würzburg, 97070 Würzburg, Germany.
Lauriano C M
Klose K E
Kapfhammer D
Kraiss A
Reidl J
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/IAI.69.1.435-445.2001
Published
2001-01-00
Pages
435-45
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC97901
Subset
IM
Grants
NIAID NIH HHS · R01 AI043486 · United States
NIAID NIH HHS · AI43486 · United States
External Links
PubMed source DOI: 10.1128/IAI.69.1.435-445.2001 PMC Full Text View journal details
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