Haemophilus ducreyi requires the flp gene cluster for microcolony formation in vitro

Joseph R. Nika, Jo L. Latimer, Christine K. Ward, Robert J. Blick, Nikki J. Wagner, Leslie D. Cope, Gregory G. Mahairas, Robert S. Munson, Eric J. Hansen

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Abstract

Haemophilus ducreyi, the etiologic agent of chancroid, has been shown to form microcolonies when cultured in the presence of human foreskin fibroblasts. We identified a 15-gene cluster in H. ducreyi that encoded predicted protein products with significant homology to those encoded by the tad (for tight adhesion) locus in Actinobacillus actinomycetemcomitans that is involved in the production of fimbriae by this periodontal pathogen. The first three open reading frames in this H. ducreyi gene cluster encoded predicted proteins with a high degree of identity to the Flp (fimbria-like protein) encoded by the first open reading frame of the tad locus; this 15-gene cluster in H. ducreyi was designated flp. RT-PCR analysis indicated that the H. ducreyi flp gene cluster was likely to be a polycistronic operon. Mutations within the flp gene cluster resulted in an inability to form microcolonies in the presence of human foreskin fibroblasts. In addition, the same mutants were defective in the ability to attach to both plastic and human foreskin fibroblasts in vitro. An H. ducreyi mutant with an inactivated tadA gene exhibited a small decrease in virulence in the temperature-dependent rabbit model for experimental chancroid, whereas another H. ducreyi mutant with inactivated flp-1 and flp-2 genes was as virulent as the wild-type parent strain. These results indicate that the flp gene cluster is essential for microcolony formation by H. ducreyi, whereas this phenotypic trait is not linked to the virulence potential of the pathogen, at least in this animal model of infection.

Original languageEnglish (US)
Pages (from-to)2965-2975
Number of pages11
JournalInfection and Immunity
Volume70
Issue number6
DOIs
StatePublished - 2002

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Haemophilus ducreyi
Multigene Family
Foreskin
Chancroid
Fibroblasts
Open Reading Frames
Virulence
Fimbriae Proteins
Aggregatibacter actinomycetemcomitans
In Vitro Techniques
Operon
Genes
Plastics
Proteins
Theoretical Models
Animal Models
Rabbits
Polymerase Chain Reaction
Mutation

ASJC Scopus subject areas

  • Immunology

Cite this

Nika, J. R., Latimer, J. L., Ward, C. K., Blick, R. J., Wagner, N. J., Cope, L. D., ... Hansen, E. J. (2002). Haemophilus ducreyi requires the flp gene cluster for microcolony formation in vitro. Infection and Immunity, 70(6), 2965-2975. https://doi.org/10.1128/IAI.70.6.2965-2975.2002

Haemophilus ducreyi requires the flp gene cluster for microcolony formation in vitro. / Nika, Joseph R.; Latimer, Jo L.; Ward, Christine K.; Blick, Robert J.; Wagner, Nikki J.; Cope, Leslie D.; Mahairas, Gregory G.; Munson, Robert S.; Hansen, Eric J.

In: Infection and Immunity, Vol. 70, No. 6, 2002, p. 2965-2975.

Research output: Contribution to journalArticle

Nika, JR, Latimer, JL, Ward, CK, Blick, RJ, Wagner, NJ, Cope, LD, Mahairas, GG, Munson, RS & Hansen, EJ 2002, 'Haemophilus ducreyi requires the flp gene cluster for microcolony formation in vitro', Infection and Immunity, vol. 70, no. 6, pp. 2965-2975. https://doi.org/10.1128/IAI.70.6.2965-2975.2002
Nika, Joseph R. ; Latimer, Jo L. ; Ward, Christine K. ; Blick, Robert J. ; Wagner, Nikki J. ; Cope, Leslie D. ; Mahairas, Gregory G. ; Munson, Robert S. ; Hansen, Eric J. / Haemophilus ducreyi requires the flp gene cluster for microcolony formation in vitro. In: Infection and Immunity. 2002 ; Vol. 70, No. 6. pp. 2965-2975.
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abstract = "Haemophilus ducreyi, the etiologic agent of chancroid, has been shown to form microcolonies when cultured in the presence of human foreskin fibroblasts. We identified a 15-gene cluster in H. ducreyi that encoded predicted protein products with significant homology to those encoded by the tad (for tight adhesion) locus in Actinobacillus actinomycetemcomitans that is involved in the production of fimbriae by this periodontal pathogen. The first three open reading frames in this H. ducreyi gene cluster encoded predicted proteins with a high degree of identity to the Flp (fimbria-like protein) encoded by the first open reading frame of the tad locus; this 15-gene cluster in H. ducreyi was designated flp. RT-PCR analysis indicated that the H. ducreyi flp gene cluster was likely to be a polycistronic operon. Mutations within the flp gene cluster resulted in an inability to form microcolonies in the presence of human foreskin fibroblasts. In addition, the same mutants were defective in the ability to attach to both plastic and human foreskin fibroblasts in vitro. An H. ducreyi mutant with an inactivated tadA gene exhibited a small decrease in virulence in the temperature-dependent rabbit model for experimental chancroid, whereas another H. ducreyi mutant with inactivated flp-1 and flp-2 genes was as virulent as the wild-type parent strain. These results indicate that the flp gene cluster is essential for microcolony formation by H. ducreyi, whereas this phenotypic trait is not linked to the virulence potential of the pathogen, at least in this animal model of infection.",
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