Autophosphorylation and phosphotransfer in the Bordetella pertussis BvgAS signal transduction cascade.

Uhl, Michaël; Miller, Jeff F.

doi:10.1073/pnas.91.3.1163

Cited by 133 publications

(137 citation statements)

References 26 publications

(23 reference statements)

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“…The N-terminal periplasmic domain of BvgS is involved in signal recognition; via phosphorylation, it transduces the signals into the transcriptional regulation network (Uhl & Miller, 1994). Evidence that BvgS is involved in signalling events is provided by the finding of a class of mutations that make the system insensitive to environmental regulation (Miller et al, 1992;Manetti e t al., 1994;Goyard e t al., 1994).…”

Section: A P R U G N O L a A N D O T H E R Smentioning

confidence: 99%

Response of the bvg regulon of Bordetella pertussis to different temperatures and short-term temperature shifts

Prugnola

Aricò²,

Manetti³

et al. 1995

Microbiology

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Bordetella pertussis produces a number of virulence factors whose expression is coordinately regulated by the bvgA3 locus. Transcription of virulence genes is repressed by environmental factors such as low temperature (25 "C) and chemical stimuli. Temperature shift of bacterial cultures from 25 "C to 37 "C activates two classes of bvg-regulated virulence genes: the early genes, which are activated within 10 min, and late genes, which require 2 4 h for activation. During the interval between the activation of the early and late genes, the intracellular concentration of BvgA increases 50-fold. It has been proposed that this increased concentration may be required for the activation of the late genes. Here we have analysed the response of the bvg locus to intermediate temperatures and to repeated temperature shifts. Temperature shifts of B.pertussis cultures from 22 "C to 28 "C, 32 "C or 35 "C resulted in the synthesis of low, intermediate, and high amounts of BvgA. This implied that the intracellular concentration of BvgA is temperature-dependent. We have also observed that the amount of virulence factors produced correlates with the BvgA concentration. When bacteria grown at 37 "C were shifted to 22 "C, transcription from the adenylate cyclase toxin haemolysis promoter (PAC) was repressed after 30 min, while transcription from the bvg (P,) and filamentous haemagglutinin (PFHA) promoters was repressed after 2 h. During this time, the amount of BvgA did not decrease. A subsequent temperature shift from 22 "C to 37 "C induced transcription from the P, and P, , promoters after 10 min and transcription from the PAC promoter after 20 min. This result shows that in the presence of a high concentration of BvgA, the time lag between temperature shift and late promoter transcription is reduced from 2 4 h to 20 min. The above data support the proposal that the concentration of BvgA plays a role in activating expression of the late genes.

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Section: A P R U G N O L a A N D O T H E R Smentioning

confidence: 99%

Response of the bvg regulon of Bordetella pertussis to different temperatures and short-term temperature shifts

Prugnola

Aricò²,

Manetti³

et al. 1995

Microbiology

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“…In the Evg system of E. coli and the Bvg system of Bordetella pertussis, three signal transduction modules (the transmitter, the receiver and the C-terminal domain containing the HPt motif) are combined in a single protein, the EvgS or BvgS sensory protein, whereas the corresponding effector molecules are typical response regulators (EvgA and BvgA) belonging to the FixJ family with an N-terminal receiver and a C-terminal DNA-binding domain Stibitz and Yang, 1991;Boucher et al, 1994;Uhl and Miller, 1994;Utsumi et al, 1994). Both systems show very significant sequence similarities even outside of the conserved signalling modules and, accordingly, seem to respond to similar external stimuli Utsumi et al, 1994;.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, it has been shown that, similar to the Bacillus and Saccharomyces systems, a complex phosphorelay occurs that involves the autophosphorylation of the BvgS transmitter (His-729) and subsequent phosphorylation of Asp-1023 in its receiver and of His-1172 in its HPt domain before the phosphorylation of BvgA (Uhl and Miller, 1994;1996a). The C-terminal receiver and HPt BvgS domains are essential for signal transduction (Uhl and Miller, 1994;Beier et al, 1995;. The phosphorelay very probably involves dimerization (or oligomerization) of BvgS (Beier et al, 1995;.…”

Section: Introductionmentioning

confidence: 99%

Specificity of the BvgAS and EvgAS phosphorelay is mediated by the C‐terminal HPt domains of the sensor proteins

Perraud

Kimmel

Weiss

et al. 1998

Molecular Microbiology

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SummaryDespite the presence of highly conserved signalling modules, significant cross-communication between different two-component systems has only rarely been observed. Domain swapping and the characterization of liberated signalling modules enabled us to characterize in vitro the protein domains that mediate specificity and are responsible for the high fidelity in the phosphorelay of the unorthodox Bvg and Evg twocomponent systems. Under equimolar conditions, significant in vitro phosphorylation of purified BvgA and EvgA proteins was only obtained by their histidine kinases, BvgS and EvgS respectively. One hybrid histidine kinase consisting of the BvgS transmitter and HPt domains and of the EvgS receiver domain (BvgS-TO-EvgS-R) was able to phosphorylate BvgA but not EvgA. In contrast, the hybrid protein consisting of the BvgS transmitter and the EvgS receiver and HPt domains (BvgS-T-EvgS-RO) was unable to phosphorylate BvgA but efficiently phosphorylated EvgA. These results demonstrate that the C-terminal HPt domains of the sensor proteins endow the unorthodox twocomponent systems with a high specificity for the corresponding regulator protein. In the case of the response regulators, the receiver but not the output domains contribute to the specific interaction with the histidine kinases, because a hybrid protein consisting of the EvgA receiver and the BvgA output domain could only be phosphorylated by the EvgS protein.

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“…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)(51)(52)(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.…”

mentioning

confidence: 99%

“…The BvgS periplasmic region has been postulated to sense signals and transmit information to the cytoplasmic domains via the BvgS linker, which lies adjacent to the second transmembrane domain (32). The cytoplasmic portion of BvgS effects the enzymatic steps of signal transduction and can autophosphorylate in vitro with the ␥-phosphate from ATP and transfer a phosphoryl group to BvgA (53,54). BvgA contains an N-terminal receiver domain and a C-terminal helix-turn-helix DNA binding motif (4), and phosphorylation of BvgA has been shown to increase its affinity for specific target sequences (7).…”

mentioning

confidence: 99%

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

Uhl

Miller

1995

J Bacteriol

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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...

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Autophosphorylation and phosphotransfer in the Bordetella pertussis BvgAS signal transduction cascade.

Cited by 133 publications

References 26 publications

Response of the bvg regulon of Bordetella pertussis to different temperatures and short-term temperature shifts

Response of the bvg regulon of Bordetella pertussis to different temperatures and short-term temperature shifts

Specificity of the BvgAS and EvgAS phosphorelay is mediated by the C‐terminal HPt domains of the sensor proteins

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

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