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

BMC Microbiology

Rita Tamayo¹  Courtney Whitaker¹  Benjamin Mudrak¹  Ankunda Kariisa¹  Robert W McKee¹ 

[1] Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, 125 Mason Farm Rd, Chapel Hill, CB# 7290, NC, USA

关键词: HD-GYP;    Virulence;    Motility;    Biofilm;    Phosphodiesterase;    Cyclic diguanylate;    Second messenger;   

Others  :  1137742
DOI  :  10.1186/s12866-014-0272-9

 received in 2014-06-18, accepted in 2014-10-16,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 McKee et al.; licensee BioMed Central Ltd.

【 图 表 】

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