A systematic analysis of the in vitro and in vivo functions of the HD-GYP domain proteins of Vibrio cholerae Public Deposited

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Creator
  • Mudrak, Benjamin
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Kariisa, Ankunda T
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Tamayo, Rita
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • McKee, Robert W
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Whitaker, Courtney
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
Abstract
  • Abstract Background The second messenger cyclic diguanylate (c-di-GMP) plays a central role in bacterial adaptation to extracellular stimuli, controlling processes such as motility, biofilm development, cell development and, in some pathogens, virulence. The intracellular level of c-di-GMP is controlled by the complementary activities of diguanylate cyclases containing a GGDEF domain and two classes of c-di-GMP phosphodiesterases containing an EAL or HD-GYP hydrolytic domain. Compared to the GGDEF and EAL domains, the functions of HD-GYP domain family proteins are poorly characterized. The human diarrheal pathogen Vibrio cholerae encodes nine putative HD-GYP domain proteins. To determine the contributions of HD-GYP domain proteins to c-di-GMP signaling in V. cholerae, we systematically analyzed the enzymatic functionality of each protein and their involvement in processes known to be regulated by c-di-GMP: motility, biofilm development and virulence. Results Complementary in vitro and in vivo experiments showed that four HD-GYP domain proteins are active c-di-GMP phosphodiesterases: VC1295, VC1348, VCA0210 and VCA0681. Mutation of individual HD-GYP domain genes, as well as combinatorial mutations of multiple HD-GYP domain genes, had no effect on motility or biofilm formation of V. cholerae under the conditions tested. Furthermore, no single HD-GYP domain gene affected intestinal colonization by V. cholerae in an infant mouse model. However, inactivation of multiple HD-GYP domain genes, including the four encoding functional phosphodiesterases, significantly attenuated colonization. Conclusions These results indicate that the HD-GYP family of c-di-GMP phosphodiesterases impacts signaling by this second messenger during infection. Altogether, this work greatly furthers the understanding of this important family of c-di-GMP metabolic enzymes and demonstrates a role for HD-GYP domain proteins in the virulence of V. cholerae.
Date of publication
Identifier
  • doi:10.1186/s12866-014-0272-9
Resource type
  • Article
Rights statement
  • In Copyright
Rights holder
  • Robert W McKee et al.; licensee BioMed Central Ltd.
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Journal title
  • BMC Microbiology
Journal volume
  • 14
Journal issue
  • 1
Page start
  • 272
Language
  • English
Is the article or chapter peer-reviewed?
  • Yes
ISSN
  • 1471-2180
Bibliographic citation
  • BMC Microbiology. 2014 Oct 25;14(1):272
Access
  • Open Access
Publisher
  • BioMed Central Ltd
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