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PMID: 7927748 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Cloning and sequencing of a Bordetella pertussis serum resistance locus.

Infection and immunity ·Vol. 62 ·No. 11 ·1994-11-00 ·Pages 4727-38

Fernandez RC, Weiss AA

Abstract

We have characterized a new virulence factor in Bordetella pertussis: serum resistance. Compared with Escherichia coli HB101, wild-type B. pertussis was relatively resistant to classical-pathway, complement-dependent killing by normal human serum. However, a mutant of B. pertussis (BPM2041) which is less virulent in mice and which has Tn5 lac inserted in a previously uncharacterized bvg-regulated gene was found to be at least 10-fold more susceptible to serum killing than the wild type. We have named this locus brk, for Bordetella resistance to killing. We have cloned and sequenced the brk locus, and it encodes two divergently transcribed open reading frames (ORFs), termed BrkA and BrkB. Both ORFs are necessary for serum resistance. Within the 300 bases which separate the two ORFs and upstream of each ORF are putative sites for BvgA binding. BrkA shows 29% identity to pertactin and has two RGD motifs in addition to a conserved proteolytic processing site and an outer membrane targeting signal. Like pertactin, BrkA is involved in adherence and invasion. Despite the similarities, a pertactin mutant was found to be not as sensitive to serum killing as the BrkA or BrkB mutants. BrkB is similar to ORFs in E. coli and Mycobacterium leprae and displays domains of homology to various transporters. On the basis of its hydropathy profile, BrkB is predicted to be a cytoplasmic membrane protein. By Southern blot, brk sequences were found in Bordetella bronchiseptica and Bordetella parapertussis but not in Bordetella avium.

Related Genes
brkA brkB cpn60
MeSH Terms
Amino Acid Sequence Bacterial Adhesion Bacterial Outer Membrane Proteins/chemistry,genetics Bacterial Proteins Base Sequence Blood Bactericidal Activity Bordetella pertussis/genetics,pathogenicity Chaperonin 60/genetics Chaperonins/genetics Cloning, Molecular Complement Pathway, Classical Genes, Bacterial Humans In Vitro Techniques Membrane Proteins/chemistry,genetics Molecular Sequence Data Molecular Weight Mutagenesis, Insertional Restriction Mapping Species Specificity Virulence Factors, Bordetella
Chemicals
Bacterial Outer Membrane Proteins Bacterial Proteins BrkA protein, Bordetella pertussis BrkB protein, Bordetella pertussis Chaperonin 60 Membrane Proteins Virulence Factors, Bordetella pertactin Chaperonins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fernandez R C
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, Ohio 45267.
Weiss A A
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.62.11.4727-4738.1994
Published
1994-11-00
Pages
4727-38
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC303180
Subset
IM
Grants
NIAID NIH HHS · AI-23695 · United States
Databases
GENBANK
U12276
External Links
PubMed source DOI: 10.1128/iai.62.11.4727-4738.1994 PMC Full Text View journal details
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