Mutation of the htrB gene in a virulent Salmonella typhimurium strain by intergeneric transduction: strain construction and phenotypic characterization

Melvin G Sunshine; Bradford W Gibson; Jeffrey J Engstrom; Wade A Nichols; Bradley D Jones; Michael A Apicella

doi:10.1128/jb.179.17.5521-5533.1997

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Mutation of the htrB gene in a virulent Salmonella typhimurium strain by intergeneric transduction: strain construction and phenotypic characterization

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Mutation of the htrB gene in a virulent Salmonella typhimurium strain by intergeneric transduction: strain construction and phenotypic characterization

Melvin G Sunshine, Bradford W Gibson, Jeffrey J Engstrom, Wade A Nichols, Bradley D Jones and Michael A Apicella

Journal of bacteriology, Vol.179(17), pp.5521-5533

09/1997

DOI: 10.1128/jb.179.17.5521-5533.1997

PMCID: PMC179425

PMID: 9287009

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Abstract

The htrB gene product of Haemophilus influenzae contributes to the toxicity of the lipooligosaccharide. The htrB gene encodes a 2-keto-3-deoxyoctulosonic acid-dependent acyltransferase which is responsible for myristic acid substitutions at the hydroxy moiety of lipid A beta-hydroxymyristic acid. Mass spectroscopic analysis has demonstrated that lipid A from an H. influenzae htrB mutant is predominantly tetraacyl and similar in structure to lipid IV(A), which has been shown to be nontoxic in animal models. We sought to construct a Salmonella typhimurium htrB mutant in order to investigate the contribution of htrB to virulence in a well-defined murine typhoid model of animal pathogenesis. To this end, an r- m+ galE mutS recD strain of S. typhimurium was constructed (MGS-7) and used in inter- and intrastrain transduction experiments with both coliphage P1 and Salmonella phage P22. The Escherichia coli htrB gene containing a mini-Tn10 insertion was transduced from E. coli MLK217 into S. typhimurium MGS-7 via phage P1 and subsequently via phage P22 into the virulent Salmonella strain SL1344. All S. typhimurium transductants showed phenotypes similar to those described for the E. coli htrB mutant. Mass spectrometric analysis of the crude lipid A fraction from the lipopolysaccharide of the S. typhimurium htrB mutant strain showed that for the dominant hexaacyl form, a lauric acid moiety was lost at one position on the lipid A and a palmitic acid moiety was added at another position; for the less abundant heptaacyl species, the lauric acid was replaced with palmitoleic acid.

Details

Title: Subtitle: Mutation of the htrB gene in a virulent Salmonella typhimurium strain by intergeneric transduction: strain construction and phenotypic characterization
Creators: Melvin G Sunshine - Department of Microbiology, The University of Iowa, Iowa City 52242, USA
Bradford W Gibson - Department of Microbiology, The University of Iowa, Iowa City 52242, USA
Jeffrey J Engstrom - Department of Microbiology, The University of Iowa, Iowa City 52242, USA
Wade A Nichols - Department of Microbiology, The University of Iowa, Iowa City 52242, USA
Bradley D Jones - Department of Microbiology, The University of Iowa, Iowa City 52242, USA
Michael A Apicella - Department of Microbiology, The University of Iowa, Iowa City 52242, USA
Resource Type: Journal article
Publication Details: Journal of bacteriology, Vol.179(17), pp.5521-5533
DOI: 10.1128/jb.179.17.5521-5533.1997
PMID: 9287009
PMCID: PMC179425
NLM abbreviation: J Bacteriol
ISSN: 0021-9193
eISSN: 1098-5530
Language: English
Date published: 09/1997
Academic Unit: Microbiology and Immunology; Internal Medicine
Record Identifier: 9984083866602771

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