title: Molecular characterization of catalase from Bordetella pertussis: identification of the katA promoter in an upstream insertion sequence.
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.
authors: Department of Microbiology and Immunology, University of Arizona, Tucson 85724.
authors: D
authors: D
authors: DeShazer
authors: G E
authors: GE
authors: Wood
authors: R L
authors: RL
authors: Friedman
mesh_terms: D001483:Base Sequence; D001886:Bordetella pertussis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D002374:Catalase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D017931:DNA Primers; D004251:DNA Transposable Elements; D017353:Gene Deletion; D005798:Genes, Bacterial; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007700:Kinetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000382:microbiology; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D015183:Restriction Mapping; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers; D004251:DNA Transposable Elements; D012333:RNA, Messenger; D006861:Hydrogen Peroxide; D002374:Catalase