PMID:16597943

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Citation

Domka, J, Lee, J and Wood, TK (2006) YliH (BssR) and YceP (BssS) regulate Escherichia coli K-12 biofilm formation by influencing cell signaling.Appl. Environ. Microbiol. 72:2449-59

Abstract

We previously discovered that yliH and yceP are induced in Escherichia coli biofilms (D. Ren, L. A. Bedzyk, S. M. Thomas, R. W. Ye, and T. K. Wood, Appl. Microbiol. Biotechnol. 64:515-524, 2004). Here, it is shown that deletion of yceP (b1060) and yliH (b0836) increases biofilm formation in continuous-flow chambers with minimal glucose medium by increasing biofilm mass (240- to 290-fold), surface coverage (16- to 31-fold), and mean thickness (2,800-fold). To determine the genetic basis of the increase in biofilm formation, we examined the differential gene expression profile in biofilms for both the mutants relative to the wild-type strain in rich medium with glucose and found that 372 to 882 genes were induced and that 76 to 337 were repressed consistently >2-fold (P < or = 0.05). The increase in biofilm formation was related to differential expression of genes related to stress response (8 to 64 genes) for both mutants, including rpoS and sdiA. More importantly, 42 to 130 genes related to autoinducer 2 cell signaling were also differentially expressed, including gadAB and flgBCEGHIJLMN, as well as signaling through indole, since 17 to 26 indole-related genes were differentially expressed, including phoAER, gltBD, mtr (encodes protein for indole import), and acrEF (encodes proteins for indole export). Increased biofilm formation in the yliH and yceP mutants in LB supplemented with 0.2% glucose (LB glu) occurred through a reduction in extracellular and intracellular indole concentrations in both mutants (50- to 140-fold), and the addition of indole to the culture restored the wild-type biofilm phenotype; hence, indole represses biofilms. Additionally, both mutants regulate biofilms through quorum sensing, since deletion of either yliH or yceP increased extracellular autoinducer 2 concentrations 50-fold when grown in complex medium (most notably in the stationary phase). Both proteins are involved in motility regulation, since YliH (127 amino acids) and YceP (84 amino acids) repressed motility two to sevenfold (P < or = 0.05) in LB, and YceP repressed motility sevenfold (P < or = 0.05) in LB glu. Heightened motility in the yceP mutant occurred, due to increased transcription of the flagella and motility loci, including fliC, motA, and qseB (3- to 86-fold). We propose new names for these two loci: bssR for yliH and bssS for yceP, based on the phrase "regulator of biofilm through signal secretion."

Links

PubMed Online version:10.1128/AEM.72.4.2449-2459.2006

Keywords

Biofilms; Culture Media; Escherichia coli K12; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Homoserine; Indoles; Lactones; Movement; Mutation; Oligonucleotide Array Sequence Analysis; Sequence Analysis, DNA; Signal Transduction

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:
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: bssS
  • Genotype of Experimental Strain : bssS(del)
  • Reference Condition:

OMP:0000041

increased cell size

ECO:0000176

mutant visible phenotype analysis

Mutation: table 2.

  • increase biofilm mass
  • increase biofilm mass
  • increase surface coverage
  • increase mean thickness

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain:
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: bssR
  • Genotype of Experimental Strain : bssR(del)
  • Reference Condition:

OMP:0000041

increased cell size

ECO:0000176

mutant visible phenotype analysis

Mutant: table 2.

  • increase biofilm formation
  • increase biofilm mass
  • increase surface coverage
  • increase mean thickness

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain:
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: BssR
  • Genotype of Experimental Strain : BssR(del)
  • Reference Condition:

OMP:0000045

increased motility

ECO:0000176

mutant visible phenotype analysis

Deletion mutation led to an increase in motility, relative to the wild type in LB medium, fig 3A. Except when glucose was added where motility was inhibited.


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