Characterization of a WaaF (RfaF) homolog expressed by haemophilus ducreyi

Beth A. Bauer, Sheryl R. Lumbley, Eric J. Hansen

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Haemophilus ducreyi lipooligosaccharide (LOS) is capable of inducing an inflammatory response in skin (A. A. Campagnari, L. M. Wild, G. Griffiths, R. J. Karalus, M. A. Wirth, and S. M. Spinola, Infect. Immun. 59:2601-2608, 1991) and likely contributes to the virulence of this sexually transmitted pathogen (B. A. Bauer, M. K. Stevens, and E. J. Hansen, Infect. Immun. 68:4290-4298, 1998). An open reading frame in H. ducreyi 35000 was found to encode a predicted protein that was 59% identical to the protein product of the rfaF (waaF) gene of Salmonella typhimurium. The H. ducreyi waaF gene was able to complement an S. typhimurium rfaF (waaF) mutant, a result which confirmed the identity of this gene. In contrast to the rfaF (waaF) gene of enteric bacteria, the H. ducreyi waaF gene was not located adjacent to other genes involved in lipopolysaccharide expression. Inactivation of the H. ducreyi waaF gene by insertion mutagenesis resulted in expression of a LOS that migrated much faster than wild-type LOS in sodium dodecyl sulfate- polyacrylamide gel electrophoresis. The LOS of this mutant also did not bind a monoclonal antibody directed against a cell surface-exposed epitope of wild-type H. ducreyi LOS. Testing of the wild-type H. ducreyi strain and its isogenic waaF mutant in the temperature-dependent rabbit model for dermal lesion production by H. ducreyi revealed that this waaF mutant was less virulent than the wild-type parent strain. Complementation of the H. ducreyi waaF mutant with the wild-type H. ducreyi waaF gene resulted in expression of both wild-type LOS and wild-type virulence by this mutant.

Original languageEnglish (US)
Pages (from-to)899-907
Number of pages9
JournalInfection and Immunity
Volume67
Issue number2
StatePublished - 1999

Fingerprint

Haemophilus ducreyi
Genes
Salmonella typhimurium
Virulence
Skin
Insertional Mutagenesis
Enterobacteriaceae
Mutagenesis
Sodium Dodecyl Sulfate
Open Reading Frames
Lipopolysaccharides
lipid-linked oligosaccharides
Epitopes
Polyacrylamide Gel Electrophoresis
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Characterization of a WaaF (RfaF) homolog expressed by haemophilus ducreyi. / Bauer, Beth A.; Lumbley, Sheryl R.; Hansen, Eric J.

In: Infection and Immunity, Vol. 67, No. 2, 1999, p. 899-907.

Research output: Contribution to journalArticle

Bauer, Beth A. ; Lumbley, Sheryl R. ; Hansen, Eric J. / Characterization of a WaaF (RfaF) homolog expressed by haemophilus ducreyi. In: Infection and Immunity. 1999 ; Vol. 67, No. 2. pp. 899-907.
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