Molecular characterization of catalase from Bordetella pertussis: Identification of the katA promoter in an upstream insertion sequence

David DeShazer, Gwendolyn E. Wood, Richard L Friedman

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

In this report we evaluate the role of catalase in the survival of Bordetella pertussis within human polymorphonuclear leukocytes (PMNs). Crude extracts of B. pertussis exhibited a single catalase activity when subjected to non-denaturing polyacrylamide gel electrophoresis and assayed far catalase activity. A plasmid containing B. pertussis katA was identified by complementation of UM255, a catalase-deficient strain of Escherichia coli. The nucleotide sequence of katA predicts a 55 kDa protein that shares homology with a class of haem-containing catalases found in both eubacteria and eukaryotes. Analysis of the nucleotide sequence upstream of katA revealed the presence of a copy of IS481, a B. pertussis-specific insertion sequence. The start site of transcription of katA was mapped to a T residue in IS481 by primer extension analysis performed with B. pertussis RNA and a katA-specific primer. A catalase-deficient strain of B. pertussis, DD900, was constructed by gene replacement. DD900 was more sensitive to killing by 1 and 5 mM H2O3 than the parental strain, BP339. However, there was no difference in the ability of DD900 and BP339 to survive for 2h in human PMNs. This suggests that catalase plays no significant role in the survival of B. pertussis within PMNs.

Original languageEnglish (US)
Pages (from-to)123-130
Number of pages8
JournalMolecular Microbiology
Volume14
Issue number1
DOIs
StatePublished - Oct 1994

Fingerprint

Bordetella pertussis
Insertional Mutagenesis
Catalase
Native Polyacrylamide Gel Electrophoresis
Survival
Transcription Initiation Site
Eukaryota
Complex Mixtures
Heme
Neutrophils
Plasmids
RNA
Escherichia coli
Bacteria
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Molecular characterization of catalase from Bordetella pertussis : Identification of the katA promoter in an upstream insertion sequence. / DeShazer, David; Wood, Gwendolyn E.; Friedman, Richard L.

In: Molecular Microbiology, Vol. 14, No. 1, 10.1994, p. 123-130.

Research output: Contribution to journalArticle

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