Extracytoplasmic adenylate cyclase of Bordetella pertussis.

Hewlett, Erik L.; Urban, Milan; Manclark, C R; Wolff, J.

doi:10.1073/pnas.73.6.1926

Cited by 140 publications

(85 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Murine monoclonal antibodies used for detection were BPH2, specific for serotype 2 fimbrial subunit (4a). BPE3, specific for pertactin (5); and 9D4, specific for adenylate cyclase toxin (13). BPH2 and BPE3 were kindly provided by LacZ' and may be Hly-as well.…”

Section: Methodsmentioning

confidence: 99%

Mutations in the bvgA gene of Bordetella pertussis that differentially affect regulation of virulence determinants

Stibitz

1994

J Bacteriol

View full text Add to dashboard Cite

By using chemical mutagenesis and genetic mapping, a search was undertaken for previously undescribed genes which may be involved in different regulatory mechanisms governing different virulence factors of Bordetella pertussis. Previous studies have shown that the fha locus encoding filamentous hemagglutinin is regulated directly by the bvgAS two component system, while regulation of ptx encoding pertussis toxin is less direct or occurs by a different mechanism. With a strain containing gene fusions to each of these regulated loci, screening was done for mutations which were defective for ptx expression but maintained normal or nearly normal levels of fha expression. Two mutations which had such a phenotype and were also deficient in adenylate cyclase toxin/hemolysin expression were found and characterized more fully. Both were found to affect residues in the C-terminal portion of the BvgA response regulator protein, a domain which shares sequence similarity with a family of regulatory proteins including FixJ, UhpA, MalT, RcsA, RcsB, and LuxR. The residues affected are within a region which, by extension from studies on the LuxR protein, may be involved in transcriptional activation.

show abstract

Section: Methodsmentioning

confidence: 99%

Mutations in the bvgA gene of Bordetella pertussis that differentially affect regulation of virulence determinants

Stibitz

1994

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…Our results demonstrate Bvg-dependent activation of a ptxA-lacZYA fusion in E. coli and indicate that bvg is the only Bordetella locus required for ptxA activation in this heterologous system. Bordetella pertussis, the causative agent of whooping cough, synthesizes toxins and adhesins which appear to aid the bacterium in colonization and multiplication (15,23,28,38,56,59). Expression of toxins (pertussis toxin, adenylate cyclasehemolysin, and dermonecrotic toxin) and adhesins (filamentous hemagglutinin, fimbriae, and pertactin) is positively regulated by the products of the bvg operon (11,57,59).…”

mentioning

confidence: 99%

BvgAS is sufficient for activation of the Bordetella pertussis ptx locus in Escherichia coli

Uhl

Miller

1995

J Bacteriol

View full text Add to dashboard Cite

BvgA and BvgS, which regulate virulence gene expression in Bordetella pertussis, are members of the two-component signal transduction family. The effects of growth conditions on the ability of BvgAS to activate transcription of fhaB (encoding filamentous hemagglutinin) and ptxA (encoding the S1 subunit of pertussis toxin) were assessed in Escherichia coli by using chromosomal fhaB-lacZYA and ptxA-lacZYA fusions. Although it had previously been reported that a ptxA-lacZYA transcriptional fusion was not activated by bvgAS in E. coli (J. F. Miller, C. R. Roy, and S. Falkow, J. Bacteriol. 171:6345-6348, 1989), we now present evidence that ptxA is activated by bvgAS in E. coli in a manner that is highly dependent on the growth conditions. Higher levels of ␤-galactosidase were produced by ptxA-lacZYA in the presence of bvgAS during growth in Stainer-Scholte medium or M9 minimal salts medium with glucose than in Luria-Bertani medium. In contrast, the level of fhaB-lacZYA expression was high during growth in all media. Addition of modulating stimuli which inhibit BvgAS function eliminated expression of ptxA-lacZYA. Levels of ␤-galactosidase expressed from the ptx-lacZYA fusion correlated with growth rate and with the final optical density at 600 nm, suggesting that the lower growth rate in M9-glucose and Stainer-Scholte media was responsible for greater accumulation of ␤-galactosidase than was seen in Luria-Bertani medium. Overproduction of BvgA was not sufficient for activation of ptxA expression but was sufficient for fhaB expression. However, overproduction of a constitutive BvgA allele (bvgA-C1) or overproduction of BvgA in the presence of BvgS was able to activate ptxA. Our results demonstrate Bvg-dependent activation of a ptxA-lacZYA fusion in E. coli and indicate that bvg is the only Bordetella locus required for ptxA activation in this heterologous system. Bordetella pertussis, the causative agent of whooping cough, synthesizes toxins and adhesins which appear to aid the bacterium in colonization and multiplication (15,23,28,38,56,59). Expression of toxins (pertussis toxin, adenylate cyclasehemolysin, and dermonecrotic toxin) and adhesins (filamentous hemagglutinin, fimbriae, and pertactin) is positively regulated by the products of the bvg operon (11,57,59). In contrast to the multiple loci activated by bvg, vrg genes of B. pertussis (5,6,25), siderophore production in some Bordetella bronchiseptica strains (14), and motility genes of B. bronchiseptica (1-3) are repressed by bvg.Sequence analysis and in vitro studies have indicated that BvgA and BvgS are members of the two-component family of signal transduction proteins. BvgS is a 135-kDa sensor protein localized to the cytoplasmic membrane, and BvgA is a 23-kDa cytoplasmic response regulator (4, 7, 37, 49, 50-53). The signals sensed by BvgAS in vivo are not known, but BvgAS can be inactivated in vitro by growth at a low temperature or by addition of sulfate anion or nicotinic acid to the culture medium. This reversible inactivation of BvgAS is known as phenot...

show abstract

“…Low concentrations of extracellular cAMP after growth in C media cannot be accounted for by direct inhibition of adenylate cyclase activity by pro-C-mode salts, as most of these caused no more inhibition than did the pro-X-mode salt NaCI. The moderate inhibition caused by NaCl, Na,SO,, sodium succinate and sodium butyrate was presumably due to Na+ ions which Hewlett & Wolff (1976) reported to cause mild inhibition of adenylate cyclase activity. The inhibition caused by nicotinic acid and nicotinamide was more marked (about 60:; loss of activity).…”

Section: Discussionmentioning

confidence: 99%

“…In the present study, however, 100 'C for 10 min destroyed all detectable adenylate cyclase activity in B. pertussis intact cells. Hewlett & Wolff (1976) also reported that purified adenylate cyclase from B. putussis lost 90"" of its activity after 20 min at 56 "C. Clearly, further investigation of the effects of B. pertu.ssis adenylate cyclase on neutrophil function is needed.…”

Section: Discussionmentioning

confidence: 99%

“…Although it has not been shown conclusively that the nucleotides are taken up by the cells, preliminary experiments did show the incorporation of [3H]cAMP and [3H]dibutyryl CAMP into filtered X-mode cells after incubation of the cells in SS-X or SS-C medium at rates between 0.4-0.6 nmol min-' (mg dry wt)-'. Hewlett & Wolff (1976) reported that N a F strongly inhibited activity of purified B. pertussis adenylate cyclase. This was confirmed in the present study where 40 mM-NaF in the assay mixture inhibited adenylate cyclase activity of cell-lysate by 90%.…”

Section: Efject O J Added Cyclic Nuckotidesmentioning

confidence: 99%

See 1 more Smart Citation