Transcriptional analysis of the Pseudomonas aeruginosa exoenzyme S structural gene

Timothy L Yahr; Andrew K Hovey; Scott M Kulich; Dara W Frank

doi:10.1128/jb.177.5.1169-1178.1995

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Transcriptional analysis of the Pseudomonas aeruginosa exoenzyme S structural gene

Journal article

Open access

Peer reviewed

Transcriptional analysis of the Pseudomonas aeruginosa exoenzyme S structural gene

Timothy L Yahr, Andrew K Hovey, Scott M Kulich and Dara W Frank

Journal of bacteriology, Vol.177(5), pp.1169-1178

03/1995

DOI: 10.1128/jb.177.5.1169-1178.1995

PMCID: PMC176720

PMID: 7868588

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Abstract

The transcriptional regulation of the Pseudomonas aeruginosa exoS gene was investigated. Expression of exoS in P. aeruginosa PA103 was dependent upon growth in a low-cation environment and the presence of a functional exsA gene. Promoter fusion analysis indicated that a 285-bp PstI-NsiI fragment, located 5' of the exoS coding region, contained a functional promoter for exoS. Expression of the reporter gene was inducible in a low-cation growth environment and required a functional copy of exsA. Divergent promoters, coordinately regulated with exoS transcription, were identified within the PstI-NsiI fragment. A fusion derivative of ExsA, MALA3A2, was shown to bind directly to the PstI-NsiI probe. DNase I protection analysis demonstrated that MALA3A2 bound to the intergenic region between the postulated -35 boxes of each promoter region. Northern (RNA) blot analysis with probes internal to and upstream of exoS demonstrated that separate, coordinately regulated mRNAs were expressed in P. aeruginosa. These data suggested that a locus, coregulated with exoS transcription, was located upstream of exoS. DNA sequence analysis of the exoS upstream region revealed three open reading frames, ORF 1, ORF 2, and ORF 3. ORF 1 demonstrated significant homology to the SycE/YerA protein of Yersinia sp. SycE/YerA is postulated to function as a chaperone for the YopE cytotoxin. The loci encoding YopE and ExoS show similarities in genetic organization, protein composition, and regulation.

Amino Acid Sequence

DNA, Bacterial - metabolism

Genes, Bacterial - genetics

Bacterial Proteins - genetics

Molecular Sequence Data

Poly(ADP-ribose) Polymerases - biosynthesis

Genetic Complementation Test

Promoter Regions, Genetic - genetics

Sequence Analysis, DNA

Recombinant Fusion Proteins

DNA-Binding Proteins - metabolism

Sequence Homology, Amino Acid

Poly(ADP-ribose) Polymerases - genetics

Pseudomonas aeruginosa - genetics

Bacterial Toxins

ADP Ribose Transferases

Base Sequence

Trans-Activators - genetics

Protein Binding

Transcription, Genetic

Gene Expression Regulation, Bacterial

Bacterial Outer Membrane Proteins - genetics

Details

Title: Subtitle: Transcriptional analysis of the Pseudomonas aeruginosa exoenzyme S structural gene
Creators: Timothy L Yahr - Department of Microbiology, Medical College of Wisconsin, Milwaukee
Andrew K Hovey - Medical College of Wisconsin
Scott M Kulich - Medical College of Wisconsin
Dara W Frank - Medical College of Wisconsin
Resource Type: Journal article
Publication Details: Journal of bacteriology, Vol.177(5), pp.1169-1178
DOI: 10.1128/jb.177.5.1169-1178.1995
PMID: 7868588
PMCID: PMC176720
NLM abbreviation: J Bacteriol
ISSN: 0021-9193
eISSN: 1098-5530
Publisher: United States
Grant note: R29-AI31665 / NIAID NIH HHS
Language: English
Date published: 03/1995
Academic Unit: Microbiology and Immunology
Record Identifier: 9984002381502771

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