The Vibrio cholerae hybrid sensor kinase VieS contributes to motility and biofilm regulation by altering the cyclic diguanylate level
| Autor: | David W. Lazinski, Rita Tamayo, Andrew Camilli, Hector Martinez-Wilson, Anna D. Tischler |
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| Předmět: |
Histidine Kinase
Operon Mutant Gene-Expression 2-Component Systems Biology Microbial Communities and Interactions medicine.disease_cause Microbiology Signal-Transduction Pathways Bacterial Proteins medicine Kinase activity Phosphorylation Cyclic GMP Vibrio cholerae Molecular Biology Domain Manganese Virulence Reverse Transcriptase Polymerase Chain Reaction Protein Autophosphorylation Histidine kinase Gene Expression Regulation Bacterial Enzyme Activation Kinetics Response regulator Biochemistry Biofilms Mutagenesis Site-Directed Calcium Signal transduction Protein Kinases Escherichia-Coli Phosphorelay |
| Popis: | 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 Δ vieS mutant compared to the profound defect observed in a Δ vieA mutant. We also found that the Δ vieS strain has a lower level of vieA transcript and, similar to a Δ 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. |
| Databáze: | OpenAIRE |
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