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Journal of Bacteriology, June 2007, p. 4094-4107, Vol. 189, No. 11
0021-9193/07/$08.00+0     doi:10.1128/JB.01510-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

ScrG, a GGDEF-EAL Protein, Participates in Regulating Swarming and Sticking in Vibrio parahaemolyticus{triangledown}

Yun-Kyeong Kim and Linda L. McCarter*

Department of Microbiology, The University of Iowa, Iowa City, Iowa 52242

Received 26 September 2006/ Accepted 18 March 2007

In this work, we describe a new gene controlling lateral flagellar gene expression. The gene encodes ScrG, a protein containing GGDEF and EAL domains. This is the second GGDEF-EAL-encoding locus determined to be involved in the regulation of swarming: the first was previously characterized and named scrABC (for "swarming and capsular polysaccharide regulation"). GGDEF and EAL domain-containing proteins participate in the synthesis and degradation of the nucleotide signal cyclic di-GMP (c-di-GMP) in many bacteria. Overexpression of scrG was sufficient to induce lateral flagellar gene expression in liquid, decrease biofilm formation, decrease cps gene expression, and suppress the {Delta}scrABC phenotype. Removal of its EAL domain reversed ScrG activity, converting ScrG to an inhibitor of swarming and activator of cps expression. Overexpression of scrG decreased the intensity of a 32P-labeled nucleotide spot comigrating with c-di-GMP standard, whereas overexpression of scrG{Delta}EAL enhanced the intensity of the spot. Mutants with defects in scrG showed altered swarming and lateral flagellin production and colony morphology (but not swimming motility); furthermore, mutation of two GGDEF-EAL-encoding loci (scrG and scrABC) produced cumulative effects on swarming, lateral flagellar gene expression, lateral flagellin production and colony morphology. Mutant analysis supports the assignment of the primary in vivo activity of ScrG to acting as a phosphodiesterase. The data are consistent with a model in which multiple GGDEF-EAL proteins can influence the cellular nucleotide pool: a low concentration of c-di-GMP favors surface mobility, whereas high levels of this nucleotide promote a more adhesive Vibrio parahaemolyticus cell type.

* Corresponding author. Mailing address: Microbiology Department, The University of Iowa, Iowa City, IA 52242. Phone: (319) 335-9721. Fax: (319) 335-9721. E-mail: linda-mccarter{at}uiowa.edu

{triangledown} Published ahead of print on 30 March 2007.

Journal of Bacteriology, June 2007, p. 4094-4107, Vol. 189, No. 11
0021-9193/07/$08.00+0     doi:10.1128/JB.01510-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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