Dissection of the Functional and Structural Domains of Phosphorelay Histidine Kinase A of
            <i>Bacillus subtilis</i>

Fabret, Céline; Kanamaru, Kyoko; Stephenson, Keith; Dartois, Véronique; Perego, Marta; Hoch, James A.

doi:10.1128/jb.183.9.2795-2802.2001

Cited by 38 publications

(74 citation statements)

References 27 publications

(20 reference statements)

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“…S2 in the supplemental material). A PAS domain at the N terminus of B. subtilis KinA has a critical and indispensable role in the activity of the kinase (38,43).…”

Section: Resultsmentioning

confidence: 99%

Characterization of the Sporulation Initiation Pathway of Clostridium difficile and Its Role in Toxin Production

et al. 2009

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Clostridium difficile is responsible for significant mortality and morbidity in the hospitalized elderly. C. difficile spores are infectious and are a major factor contributing to nosocomial transmission. The Spo0A response regulator is the master regulator for sporulation initiation and can influence many other cellular processes. Using the ClosTron gene knockout system, we inactivated genes encoding Spo0A and a putative sporulation-associated sensor histidine kinase in C. difficile. Inactivation of spo0A resulted in an asporogeneous phenotype, whereas inactivation of the kinase reduced C. difficle sporulation capacity by 3.5-fold, suggesting that this kinase also has a role in sporulation initiation. Furthermore, inactivation of either spo0A or the kinase resulted in a marked defect in C. difficile toxin production. Therefore, Spo0A and the signaling pathway that modulates its activity appear to be involved in regulation of toxin synthesis in C. difficile. In addition, Spo0A was directly phosphorylated by a putative sporulation-associated kinase, supporting the hypothesis that sporulation initiation in C. difficile is controlled by a two-component signal transduction system rather than a multicomponent phosphorelay. The implications of these findings for C. difficile sporulation, virulence, and transmission are discussed.

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“…S2 in the supplemental material). A PAS domain at the N terminus of B. subtilis KinA has a critical and indispensable role in the activity of the kinase (38,43).…”

Section: Resultsmentioning

confidence: 99%

Characterization of the Sporulation Initiation Pathway of Clostridium difficile and Its Role in Toxin Production

et al. 2009

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“…Studies of the B. pertussis BvgAS and Bacillus subtilis Kin/ Spo phosphorelay systems have shown that sensor kinases, such as BvgS and Kin histidine kinases, form dimers prior to autophosphorylation (10,40). Given the similarity in domain structure between VieS and these sensor kinases, we predicted that VieS would also require dimerization for activity.…”

Section: Discussionmentioning

confidence: 99%

The Vibrio cholerae Hybrid Sensor Kinase VieS Contributes to Motility and Biofilm Regulation by Altering the Cyclic Diguanylate Level

Martinez-Wilson

Tamayo

Tischler³

et al. 2008

J Bacteriol

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Phosphorelay systems are important mediators of signal transduction during bacterial adaptation to new environments. Previously we described the vieSAB operon, encoding a putative three-protein component phosphorelay involved in regulating Vibrio cholerae virulence gene expression. At least part of the regulatory activity of VieSAB is exerted through the cyclic diguanylate (c-di-GMP)-degrading activity of the putative response regulator VieA. So far no direct evidence that VieSAB encodes a phosphorelay system exists. In addition, the role VieS plays in modulating VieA activity remains unclear. To address these questions, we expressed and purified VieA and a soluble cytoplasmic portion of VieS and used them in autophosphorylation and phosphotransfer assays. These assays showed that VieS has kinase activity in vitro and is able to selectively phosphorylate VieA. A phenotypic comparison revealed that deletion of vieS results in increased biofilm production comparable to that seen for deletion of vieA, whereas motility was decreased only slightly in the ⌬vieS mutant compared to the profound defect observed in a ⌬vieA mutant. We also found that the ⌬vieS strain has a lower level of vieA transcript and, similar to a ⌬vieA mutant, an increased intracellular level of c-di-GMP. Further analysis using site-directed vieA mutants showed that some of the phenotypes observed were due to the phosphorylation status of VieA. The evidence presented in this report is the first to link VieS and VieA biochemically and genetically, lending support to the hypothesis that these proteins function together in a signaling system.

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“…Finally, when we analyze and compile all the information gathered from all the previous related studies (Hoch, 1993;Wang et al, 2001;Eswaramoorthy et al, 2009) and our present data, we are able to propose a novel hypothetical functional model of the sensor histidine-R E T R A C T I O N kinase A, which we purposely dubbed the "C-shaped intertwined model" or C-SIM (Figure 9). Simply put, based on our results, we speculate that only the PAS-B and PAS-B' domains of both monomers interact strongly leading to a stable dimer and enabling the necessary interaction between the PAS-A and the autokinase domains for phosphate transfer.…”

Section: Kina-gfp Transcriptional Fusionsmentioning

confidence: 99%

Genetic and biochemical analyses of sensor kinase A in Bacillus subtilis sporulation

Nguele

Eswaramoorthy²,

Bhattacharya³

et al. 2010

Genet. Mol. Res.

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ABSTRACT. Temporal and spatial regulation of gene expression during endospore formation in Bacillus subtilis prompted us to investigate the molecular mechanisms that coordinate the phosphorelay. We targeted KinA for random mutagenesis. In addition, we constructed KinA-GFP transcriptional fusions for verification, via fluorescence. Four distinct types of sporulation-defective mutants were identified as inactive (no sporulation), hypoactive (low sporulation efficiency), isoactive (same efficiency as wild-type), and hyperactive (high efficiency) mutants. Surprisingly, the β-galactosidase activity of hyperactive mutants was barely greater than that of the wild-type strain; the only noticeable difference was early synthesis of KinA, which could allow them to activate Spo0A precociously, undergo sporulation earlier, and yield more spores. There was no fluorescence emission by the spore-defective mutant, which confirmed the presence of a truncated KinA (nonsense mutation) in inactive strains; other mutants harbored missense or silent mutations. We determined the nucleotide sequences of KinA mutants and found a conserved C-terminus region; more variability was observed in the N-terminus region, involving the PAS-A and PAS-C domains. We speculate that PAS-A, notwithstanding its ATPase activity, has only a minor role in KinA activity, whereas PAS-B was found to be indispensable. Our results emphasize the importance of R E T R A C T I O Ntemporal coordination of gene expression during the sporulation process and corroborate the necessity of Spo0A phosphorylation by KinA, which stimulates SpoIIG expression. We further propose a novel hypothetical model that purposely dubbed the "C-shaped intertwined model", which requires both homodimerization and spatial proximity between PAS-A and histidine H 405 of two different KinA molecules.

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Dissection of the Functional and Structural Domains of Phosphorelay Histidine Kinase A of Bacillus subtilis

Cited by 38 publications

References 27 publications

Characterization of the Sporulation Initiation Pathway of Clostridium difficile and Its Role in Toxin Production

Characterization of the Sporulation Initiation Pathway of Clostridium difficile and Its Role in Toxin Production

The Vibrio cholerae Hybrid Sensor Kinase VieS Contributes to Motility and Biofilm Regulation by Altering the Cyclic Diguanylate Level

Genetic and biochemical analyses of sensor kinase A in Bacillus subtilis sporulation

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