Molecular Characterization of Catalase From Bordetella Pertussis: Identification of the katA Promoter in an Upstream Insertion Sequence

Mol Microbiol. 1994 Oct;14(1):123-30. doi: 10.1111/j.1365-2958.1994.tb01272.x.

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 for 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 H2O2 than the parental strain, BP339. However, there was no difference in the ability of DD900 and BP339 to survive for 2 h in human PMNs. This suggests that catalase plays no significant role in the survival of B. pertussis within PMNs.

MeSH terms

  • Base Sequence
  • Bordetella pertussis / drug effects
  • Bordetella pertussis / enzymology*
  • Bordetella pertussis / genetics*
  • Catalase / biosynthesis
  • Catalase / genetics
  • Catalase / metabolism*
  • Cloning, Molecular
  • DNA Primers
  • DNA Transposable Elements*
  • Gene Deletion
  • Genes, Bacterial*
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Kinetics
  • Microbial Sensitivity Tests
  • Molecular Sequence Data
  • Neutrophils / microbiology
  • Plasmids
  • Promoter Regions, Genetic*
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis
  • Restriction Mapping
  • Transcription, Genetic

Substances

  • DNA Primers
  • DNA Transposable Elements
  • RNA, Messenger
  • Hydrogen Peroxide
  • Catalase