Cloning and characterization of btr, a Bordetella pertussis gene encoding an FNR-like transcriptional regulator

J. D. Bannan, M. J. Moran, J. I. MacInnes, G. A. Soltes, Richard L Friedman

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

To determine whether hemolytic factors other than the bifunctional hemolysin-adenylate cyclase toxin (cyclolysin) are expressed by Bordetella pertussis, a gene library was constructed from a virulent strain of B. pertussis, BP504, transformed into nonhemolytic Escherichia coli, and screened on blood agar plates. A strongly hemolytic colony which contained the plasmid pHLY1A was isolated. Nucleotide sequencing of pHLY1A revealed an open reading frame that could encode a 27-kDa protein. No similarity was detected between the deduced amino acid sequence of this open reading frame and those of any known bacterial cytolysins. However, significant homology was detected with FNR of E. coli and several other transcriptional regulators including HlyX from Actinobacillus pleuropneumoniae, which can also confer a hemolytic phenotype on E. coli. An fnr mutant of E. coli, JRG1728, could be complemented by pHLY1A. Thus, the B. pertussis transcriptional regulator- like gene and the protein which it encoded were named btr and BTR, respectively. A BTR-deficient B. pertussis strain, BJB1, was constructed. The btr::kan mutation had no effect on the expression of hemolytic activity or on phase variation. Northern (RNA) blotting revealed that btr expression was not regulated by the BvgAS two-component sensor-regulator. On the basis of sequence similarity to FNR-like transcriptional regulators and the ability to complement an anaerobically deficient E. coli strain (JRG1728) in growing anaerobically, BTR may regulate B. pertussis gene expression in response to changes in oxygen levels or to changes in the redox potential of the bacterial environment. Its role in virulence remains to be determined.

Original languageEnglish (US)
Pages (from-to)7228-7235
Number of pages8
JournalJournal of Bacteriology
Volume175
Issue number22
StatePublished - 1993

Fingerprint

Bordetella pertussis
Organism Cloning
Escherichia coli
Adenylate Cyclase Toxin
Genes
Open Reading Frames
Actinobacillus pleuropneumoniae
Hemolysin Proteins
Cytotoxins
Regulator Genes
Gene Library
Northern Blotting
Agar
Oxidation-Reduction
Virulence
Amino Acid Sequence
Proteins
Plasmids
Nucleotides
RNA

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Cloning and characterization of btr, a Bordetella pertussis gene encoding an FNR-like transcriptional regulator. / Bannan, J. D.; Moran, M. J.; MacInnes, J. I.; Soltes, G. A.; Friedman, Richard L.

In: Journal of Bacteriology, Vol. 175, No. 22, 1993, p. 7228-7235.

Research output: Contribution to journalArticle

Bannan, J. D. ; Moran, M. J. ; MacInnes, J. I. ; Soltes, G. A. ; Friedman, Richard L. / Cloning and characterization of btr, a Bordetella pertussis gene encoding an FNR-like transcriptional regulator. In: Journal of Bacteriology. 1993 ; Vol. 175, No. 22. pp. 7228-7235.
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