Cellular levels and binding of c-di-GMP control subcellular localization and activity of the Vibrio cholerae transcriptional regulator VpsT

• Published in: PLoS pathogens Vol. 8; no. 5; p. e1002719
• Main Authors: Shikuma, Nicholas J, Fong, Jiunn C N, Yildiz, Fitnat H
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01-05-2012; Public Library of Science (PLoS)
• Subjects: Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biology; Cholera; Cyclic GMP - analogs & derivatives; Cyclic GMP - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Gene Expression Regulation, Bacterial; Genetic aspects; Genetic transcription; Health aspects; Microbiology; Mutation; Phosphorus-Oxygen Lyases - genetics; Phosphorus-Oxygen Lyases - metabolism; Physiological aspects; Protein binding; Proteins; Second Messenger Systems; Signal Transduction; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription, Genetic; Vibrio; Vibrio cholerae - genetics; Vibrio cholerae - metabolism; Viral genetics; Viral Proteins - genetics; Viral Proteins - metabolism; Virulence (Microbiology); United States
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1002719

The second messenger, cyclic diguanylate (c-di-GMP), regulates diverse cellular processes in bacteria. C-di-GMP is produced by diguanylate cyclases (DGCs), degraded by phosphodiesterases (PDEs), and receptors couple c-di-GMP production to cellular responses. In many bacteria, including Vibrio cholerae, multiple DGCs and PDEs contribute to c-di-GMP signaling, and it is currently unclear whether the compartmentalization of c-di-GMP signaling components is required to mediate c-di-GMP signal transduction. In this study we show that the transcriptional regulator, VpsT, requires c-di-GMP binding for subcellular localization and activity. Only the additive deletion of five DGCs markedly decreases the localization of VpsT, while single deletions of each DGC do not impact VpsT localization. Moreover, mutations in residues required for c-di-GMP binding, c-di-GMP-stabilized dimerization and DNA binding of VpsT abrogate wild type localization and activity. VpsT does not co-localize or interact with DGCs suggesting that c-di-GMP from these DGCs diffuses to VpsT, supporting a model in which c-di-GMP acts at a distance. Furthermore, VpsT localization in a heterologous host, Escherichia coli, requires a catalytically active DGC and is enhanced by the presence of VpsT-target sequences. Our data show that c-di-GMP signaling can be executed through an additive cellular c-di-GMP level from multiple DGCs affecting the localization and activity of a c-di-GMP receptor and furthers our understanding of the mechanisms of second messenger signaling.