A 23-KILODALTON PROTEIN, DISTINCT FROM BVGA, EXPRESSED BY VIRULENT BORDETELLA-PERTUSSIS BINDS TO THE PROMOTER REGION OF VIR-REGULATED TOXIN GENES

Bordetella pertusis coordinately regulates expression of its virulence factors in response to changing environmental conditions. These factors include pertussis toxin, adenylate cyclase toxin, and the filamentous hemagglutinin (FHA). The vir (or bvg) locus has been shown genetically to be required for this coordinate regulation. We have attempted to study the biochemical basis for coordinate regulation. DNA promoter deletion studies from other laboratories have shown that two tandem 20-bp repeats - 157 to - 117 bp upstream from the pertussis toxin promoter are essential for transcription. A similar 20-bp tandem repeat was found at the same site in the upstream region of the adenylate cyclase toxin promoter but is not present in the FHA or vir promoter region. Gel retardation revealed protein from virulent strains (able to express the virulence genes) but not from avirulent strains (unable to express the virulence genes) bound to the promoter region of the pertussis toxin gene, and this binding could be abolished by competition with an excess of oligonucleotides corresponding to either tandem repeat. The protein was determined to be 23 kDa by Southwestern (DNA-protein) analysis and could bind to either 20-bp oligonucleotide from the pertussis toxin promoter and either 20-bp oligonucleotide from the adenylate cyclase toxin promoter. BvgA, a 23-kDa protein encoded in the vir locus, has been reported to bind to a 14-bp inverted repeat in the FHA promoter which is not present in the pertussis toxin or adenylate cyclase promoter. We could not demonstrate binding of BvgA to the pertussis toxin promoter region. These data suggest that we have identified a second 23-kDa protein, distinct from BvgA but regulated by the vir operon, that binds to DNA sequences required for transcription of some, but not all, vir-regulated genes.