Constitutive sensory transduction mutations in the Bordetella pertussis bvgS gene.
The products of the bvgAS locus coordinately regulate expression of the Bordetella pertussis virulence regulon in response to environmental signals. Transcription of bvgAS-activated genes is nearly eliminated by several modulating conditions, including the presence of sulfate anion or nicotinic acid and growth at low temperature. We have isolated spontaneous mutations that result in the constitutive synthesis of multiple bvg-regulated loci. Several of these mutations have been analyzed and were found to result from single-nucleotide substitutions within bvgS, in a region encoding a 161-amino-acid segment which links the transmembrane sequence with cytoplasmic domains that appear to be involved in signaling events. The effect of signal transduction mutations in Escherichia coli was determined by measuring the expression of an fhaB-lacZYA transcriptional fusion, and that in B. pertussis was determined by measuring expression of both fhaB-cat and ptxA3201-cat fusions. The constitutive mutations have little effect on fhaB-cat or fhaB-lacZYA expression in the absence of modulating signals but result in a nearly complete insensitivity to MgSO4, nicotinic acid, or growth at low temperature. Furthermore, insertion and deletion mutations in bvgS sequences encoding the periplasmic domain eliminate activity of the wild-type product, whereas constitutive mutants remain active. In B. pertussis cultures grown in Stainer-Scholte broth, expression of ptxA3201-cat differed from that of fhaB-cat in several respects. In combination with a wild-type bvgS allele, ptxA3201-cat expression required the addition of heptakis-(2,6-O-dimethyl)-beta-cyclodextrin, and this requirement was eliminated by the presence of the constitutive mutations.
Miller
J F
JF
Department of Microbiology and Immunology, School of Medicine, University of California, Los Angeles 90024.
Johnson
S A
SA
Black
W J
WJ
Beattie
D T
DT
Mekalanos
J J
JJ
Falkow
S
S
D000483:Alleles; D000595:Amino Acid Sequence; D001886:Bordetella pertussis / Q000187:drug effects / Q000235:genetics*; D002874:Chromosome Mapping; D003080:Cold Temperature; D003505:Cyclodextrins / Q000494:pharmacology; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D008278:Magnesium Sulfate / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics*; D009525:Niacin / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics*; D047392:beta-Cyclodextrins
D003505:Cyclodextrins; D011993:Recombinant Fusion Proteins; D047392:beta-Cyclodextrins; D009525:Niacin; C038119:heptakis(2,6-O-dimethyl)beta-cyclodextrin; D008278:Magnesium Sulfate