PMID:20061475

Citation	Bassis, CM and Visick, KL (2010) The cyclic-di-GMP phosphodiesterase BinA negatively regulates cellulose-containing biofilms in Vibrio fischeri. J. Bacteriol. 192:1269-78
Abstract	Bacteria produce different types of biofilms under distinct environmental conditions. Vibrio fischeri has the capacity to produce at least two distinct types of biofilms, one that relies on the symbiosis polysaccharide Syp and another that depends upon cellulose. A key regulator of biofilm formation in bacteria is the intracellular signaling molecule cyclic diguanylate (c-di-GMP). In this study, we focused on a predicted c-di-GMP phosphodiesterase encoded by the gene binA, located directly downstream of syp, a cluster of 18 genes critical for biofilm formation and the initiation of symbiotic colonization of the squid Euprymna scolopes. Disruption or deletion of binA increased biofilm formation in culture and led to increased binding of Congo red and calcofluor, which are indicators of cellulose production. Using random transposon mutagenesis, we determined that the phenotypes of the DeltabinA mutant strain could be disrupted by insertions in genes in the bacterial cellulose biosynthesis cluster (bcs), suggesting that cellulose production is negatively regulated by BinA. Replacement of critical amino acids within the conserved EAL residues of the EAL domain disrupted BinA activity, and deletion of binA increased c-di-GMP levels in the cell. Together, these data support the hypotheses that BinA functions as a phosphodiesterase and that c-di-GMP activates cellulose biosynthesis. Finally, overexpression of the syp regulator sypG induced binA expression. Thus, this work reveals a mechanism by which V. fischeri inhibits cellulose-dependent biofilm formation and suggests that the production of two different polysaccharides may be coordinated through the action of the cellulose inhibitor BinA.
Links	PubMed PMC2820850 Online version:10.1128/JB.01048-09
Keywords	Aliivibrio fischeri/physiology; Amino Acid Substitution; Animals; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Benzenesulfonates/metabolism; Biofilms/growth & development; Cellulose/metabolism; Coloring Agents/metabolism; Congo Red/metabolism; Cyclic GMP/analogs & derivatives; Cyclic GMP/metabolism; DNA Transposable Elements; Down-Regulation; Fluorescent Dyes/metabolism; Gene Knockout Techniques; Gene Order; Mutagenesis, Insertional; Mutagenesis, Site-Directed; Phosphoric Diester Hydrolases/genetics; Phosphoric Diester Hydrolases/metabolism; Staining and Labeling
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
VIBFI:CPDP	NOT	GO:0047555: 3',5'-cyclic-GMP phosphodiesterase activity	ECO:0000315:		F		Figure 3 Biofilm formation by binA syp double mutants. Mutants defective for both binA and various syp genes.	complete CACAO 8746

References

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