Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of E. coli K-12 mutants yceP, trpE, and tnaA biofilm cell 24 hr

ABSTRACT: Gene expression of (i) K-12 yceP biofilms upon addition of 600 ÂµM indole in LB glu medium, (ii) K-12 biofilms due to the trpE mutation in LB glu medium, and (iii) K-12 biofilms due to the tnaA mutation in LB medium. Experiment Overall Design: Strains:E. coli K12 BW25113 delta yceP, trpE, tnaA and wild type Experiment Overall Design: Medium:LB glu or LB Experiment Overall Design: Biofilm cells grown on glass wool Experiment Overall Design: Harvest time:24hr

ORGANISM(S): Escherichia coli

SUBMITTER: Jintae Lee

PROVIDER: E-GEOD-4562 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Indole is an inter-species biofilm signal mediated by SdiA.

Lee Jintae J Jayaraman Arul A Wood Thomas K TK

BMC microbiology 20070518

<h4>Background</h4>As a stationary phase signal, indole is secreted in large quantities into rich medium by Escherichia coli and has been shown to control several genes (e.g., astD, tnaB, gabT), multi-drug exporters, and the pathogenicity island of E. coli; however, its impact on biofilm formation has not been well-studied.<h4>Results</h4>Through a series of global transcriptome analyses, confocal microscopy, isogenic mutants, and dual-species biofilms, we show here that indole is a non-toxic si ...[more]

PMID: 17511876

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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 K-12 mutants yceP, trpE, and tnaA biofilm cell 24 hr

Publications

Indole is an inter-species biofilm signal mediated by SdiA.

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