A gene cluster involved in the utilization of both free heme and heme: hemopexin by Haemophilus influenzae type b

L. D. Cope, R. Yogev, U. Muller-Eberhard, E. J. Hansen

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

The utilization of heme bound to the serum glycoprotein hemopexin by Haemophilus influenzae type b (Hib) strain DL42 requires the presence of the 100-kDa heme:hemopexin-binding protein encoded by the hxuA gene (M. S. Hanson, S. E. Pelzel, J. Latimer, U. Muller-Eberhard, and E. J. Hansen, Proc. Natl. Acad. Sci. USA 89:1973-1977, 1992). Nucleotide sequence analysis of a 5-kb region immediately upstream from the hxuA gene revealed the presence of two genes, designated hxuC and hxuB, which encoded outer membrane proteins. The 78-kDa HxuC protein had similarity to TonB-dependent outer membrane proteins of other organisms, whereas the 60-kDa HxuB molecule most closely resembled the ShlB protein of Serratia marcescens. A set of three isogenic Hib mutants with cat cartridges inserted individually into their hxuA, hxuB, and hxuC genes was constructed. None of these mutants could utilize heme:hemopexin. The hxuC mutant was also unable to utilize low levels of free heme, whereas both the hxuA and hxuB mutants could utilize free heme. When the wild-type hxuC gene was present in trans, the hxuC mutant regained its ability to utilize low levels of free heme but still could not utilize heme:hemopexin. The hxuA mutant could utilize heme:hemopexin when a functional hxuA gene from a nontypeable H. influenzae strain was present in trans. Complementation analysis using this cloned nontypeable H. influenzae hxuA gene also indicated that the HxuB protein likely functions in the release of soluble HxuA from the Hib cell. These studies indicate that at least two and possibly three gene products are required for utilization of heme bound to hemopexin by Hib strain DL42.

Original languageEnglish (US)
Pages (from-to)2644-2653
Number of pages10
JournalJournal of Bacteriology
Volume177
Issue number10
StatePublished - 1995

Fingerprint

Hemopexin
Haemophilus influenzae type b
Multigene Family
Heme
Genes
Haemophilus influenzae
Membrane Proteins
Sequence Analysis
Glycoproteins
Proteins
Cats

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Cope, L. D., Yogev, R., Muller-Eberhard, U., & Hansen, E. J. (1995). A gene cluster involved in the utilization of both free heme and heme: hemopexin by Haemophilus influenzae type b. Journal of Bacteriology, 177(10), 2644-2653.

A gene cluster involved in the utilization of both free heme and heme : hemopexin by Haemophilus influenzae type b. / Cope, L. D.; Yogev, R.; Muller-Eberhard, U.; Hansen, E. J.

In: Journal of Bacteriology, Vol. 177, No. 10, 1995, p. 2644-2653.

Research output: Contribution to journalArticle

Cope, LD, Yogev, R, Muller-Eberhard, U & Hansen, EJ 1995, 'A gene cluster involved in the utilization of both free heme and heme: hemopexin by Haemophilus influenzae type b', Journal of Bacteriology, vol. 177, no. 10, pp. 2644-2653.
Cope, L. D. ; Yogev, R. ; Muller-Eberhard, U. ; Hansen, E. J. / A gene cluster involved in the utilization of both free heme and heme : hemopexin by Haemophilus influenzae type b. In: Journal of Bacteriology. 1995 ; Vol. 177, No. 10. pp. 2644-2653.
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AB - The utilization of heme bound to the serum glycoprotein hemopexin by Haemophilus influenzae type b (Hib) strain DL42 requires the presence of the 100-kDa heme:hemopexin-binding protein encoded by the hxuA gene (M. S. Hanson, S. E. Pelzel, J. Latimer, U. Muller-Eberhard, and E. J. Hansen, Proc. Natl. Acad. Sci. USA 89:1973-1977, 1992). Nucleotide sequence analysis of a 5-kb region immediately upstream from the hxuA gene revealed the presence of two genes, designated hxuC and hxuB, which encoded outer membrane proteins. The 78-kDa HxuC protein had similarity to TonB-dependent outer membrane proteins of other organisms, whereas the 60-kDa HxuB molecule most closely resembled the ShlB protein of Serratia marcescens. A set of three isogenic Hib mutants with cat cartridges inserted individually into their hxuA, hxuB, and hxuC genes was constructed. None of these mutants could utilize heme:hemopexin. The hxuC mutant was also unable to utilize low levels of free heme, whereas both the hxuA and hxuB mutants could utilize free heme. When the wild-type hxuC gene was present in trans, the hxuC mutant regained its ability to utilize low levels of free heme but still could not utilize heme:hemopexin. The hxuA mutant could utilize heme:hemopexin when a functional hxuA gene from a nontypeable H. influenzae strain was present in trans. Complementation analysis using this cloned nontypeable H. influenzae hxuA gene also indicated that the HxuB protein likely functions in the release of soluble HxuA from the Hib cell. These studies indicate that at least two and possibly three gene products are required for utilization of heme bound to hemopexin by Hib strain DL42.

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