Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of E.coli ydgG mutant vs. isogenic wild type in a mature biofilm

ABSTRACT: The E. coli Genechip antisense genome array was used to study the differential gene expression profile of E. coli K12 ydgG mutant compared to its isogenic wild type in a mature biofilm. Experiment Overall Design: Overnight cultures were diluted 100X into 1 L shake flasks containing 250 mL of LB + 0.2 % and 10 g of glass wool. The cells were shaken at 250 rpm and 37 C for 24 hours (final OD600=5 for the suspended cells), to form a biofilm on the glass wool, and RNA was isolated from the biofilm and Affymetrix protocol was used for preparing fragmented and labelled cDNA that was hybrizide to the chips.

ORGANISM(S): Escherichia coli

SUBMITTER: Thomas Wood

PROVIDER: E-GEOD-3514 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

YdgG (TqsA) controls biofilm formation in Escherichia coli K-12 through autoinducer 2 transport.

Herzberg Moshe M Kaye Ian K IK Peti Wolfgang W Wood Thomas K TK

Journal of bacteriology 20060101 2

YdgG is an uncharacterized protein that is induced in Escherichia coli biofilms. Here it is shown that deletion of ydgG decreased extracellular and increased intracellular concentrations of autoinducer 2 (AI-2); hence, YdgG enhances transport of AI-2. Consistent with this hypothesis, deletion of ydgG resulted in a 7,000-fold increase in biofilm thickness and 574-fold increase in biomass in flow cells. Also consistent with the hypothesis, deletion of ydgG increased cell motility by increasing tra ...[more]

PMID: 16385049

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Project description:The Escherichia coli quorum-sensing regulator, SdiA, belongs to the LuxR family of transcriptional regulators and is responsible for detecting signals from other bacteria. Previously we found that SdiA is necessary for E. coli to control its biofilm formation with indole just as SdiA is necessary for E. coli to alter its biofilm formation in the presence of N-acylhomoserine lactones (AHLs). Here we engineered SdiA by random mutagenesis to further control biofilm formation in the presence of indole and AHLs. After screening of 477?? mutants with indole and two AHLs (N-butyryl-DL-homoserine lactone, and N-(3-oxooctanoyl)-L-homoserine lactone, C6HSL), five SdiA variants were obtained that altered biofilm with and without signals of indole and AHLs. Two truncation variants (1E11 and 14C3) lacking the C-terminal DNA-binding domain of SdiA showed the reduction of biofilm formation by 5-fold and 10-fold in LB and LB glu, respectively. DNA microarrays show that the evolved SdiA (1E11) compared to wild-type SdiA influences indole synthesis genes, AI-2 uptake genes, acid-resistance genes, motility related genes, cold-shock protein genes, and several regulatory protein genes. Corroborating DNA microarrays, SdiA variants produced various amounts of indole which led to different survivals in low pH and influenced swimming motility and final cell density. Also, an AHL sensitive variant (2D10) 2-fold increased biofilm formation in the presence of C6HSL, while another variant (6B12) lowered biofilm formation in the presence of C6HSL. Hence, the results clearly showed that mutation of SdiA itself directly controls biofilm formation and SdiA variants could be further recognized by the inter-species signal AHLs. This is the first random protein engineering to control biofilm formation. Experiment Overall Design: For the microarray experiments, 10 g glass wool (Corning Glass Works, Corning, N.Y.) were used to form biofilms in 250 mL LB ( 1 mM IPTG + Cm30) in 1 L Erlenmeyer shake flasks which were inoculated with overnight cultures diluted that were 1:100. The cells were shaken at 250 rpm and 30C for 12 hours to form biofilms on the glass wool, and RNA was isolated from the the biofilm cells.

Dataset Information

Transcription profiling of E.coli ydgG mutant vs. isogenic wild type in a mature biofilm

Publications

YdgG (TqsA) controls biofilm formation in Escherichia coli K-12 through autoinducer 2 transport.

Similar Datasets

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