PMID:21181144

Citation	Sanchez-Torres, V, Hu, H and Wood, TK (2011) GGDEF proteins YeaI, YedQ, and YfiN reduce early biofilm formation and swimming motility in Escherichia coli.Appl. Microbiol. Biotechnol. 90:651-8
Abstract	The second messenger 3'-5'-cyclic diguanylic acid (c-di-GMP) promotes biofilm formation, and c-di-GMP is synthesized by diguanylate cyclases (characterized by a GGDEF domain) and degraded by phosphodiesterases. Here, we evaluated the effect of the 12 E. coli GGDEF-only proteins on biofilm formation and motility. Deletions of the genes encoding the GGDEF proteins YeaI, YedQ, YfiN, YeaJ, and YneF increased swimming motility as expected for strains with reduced c-di-GMP. Alanine substitution in the EGEVF motif of YeaI abolished its impact on swimming motility. In addition, extracellular DNA (eDNA) was increased as expected for the deletions of yeaI (tenfold), yedQ (1.8-fold), and yfiN (3.2-fold). As a result of the significantly enhanced motility, but contrary to current models of decreased biofilm formation with decreased diguanylate cyclase activity, early biofilm formation increased dramatically for the deletions of yeaI (30-fold), yedQ (12-fold), and yfiN (18-fold). Our results indicate that YeaI, YedQ, and YfiN are active diguanylate cyclases that reduce motility, eDNA, and early biofilm formation and contrary to the current paradigm, the results indicate that c-di-GMP levels should be reduced, not increased, for initial biofilm formation so c-di-GMP levels must be regulated in a temporal fashion in biofilms.
Links	PubMed Online version:10.1007/s00253-010-3074-5
Keywords	Biofilms; Cyclic GMP; Escherichia coli; Escherichia coli Proteins; Gene Deletion; Gene Expression Regulation, Bacterial; Genes, Bacterial; Mutagenesis, Site-Directed; Phosphoric Diester Hydrolases; Phosphorus-Oxygen Lyases; Protein Structure, Tertiary; Second Messenger Systems
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Phenotype of	Taxon Information	Genotype Information (if known)	Condition Information	OMP ID	OMP Term Name	ECO ID	ECO Term Name	Notes	Status
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: BW25113 NCBI Taxon ID: 679895	Genotype of Reference Strain: yeaI+ Genotype of Experimental Strain : yeaI(del)	Reference Condition: M9C, 37C, 7 h, microtiter dish wells Experimental Condition: M9C, 37C, 7h, microtiter dish wells	OMP:0007020	increased biofilm formation	ECO:0000182	in vitro culture assay evidence	Figure 1b, crystal violet biofilm assay	complete
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K12 Substrain: BW25113 NCBI Taxon ID: 679895	Genotype of Reference Strain: yeaI+ Genotype of Experimental Strain : yeaI(del)	Reference Condition: LB, 37C, 7 h Experimental Condition: LB, 37C, 7 h	OMP:0007020	increased biofilm formation	ECO:0000182	in vitro culture assay evidence	Figure 1a	complete
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K12 Substrain: BW25113 NCBI Taxon ID: 679895	Genotype of Reference Strain: yeaI+ Genotype of Experimental Strain : yeaI(del)	Reference Condition: LB, 37C, 24 h, microtiter dish wells Experimental Condition: LB, 37C, 24 h, microtiter dish wells		unaltered biofilm formation	ECO:0000182	in vitro culture assay evidence	Figure 1a	complete
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