PMID:21181144

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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

Main Points of the Paper

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Materials and Methods Used

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Phenotype Annotations

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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

Notes

References

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