Transcriptional organization of the trans-regulatory locus which controls exoenzyme S synthesis in Pseudomonas aeruginosa

T L Yahr; D W Frank

doi:10.1128/jb.176.13.3832-3838.1994

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Transcriptional organization of the trans-regulatory locus which controls exoenzyme S synthesis in Pseudomonas aeruginosa

Journal article

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Transcriptional organization of the trans-regulatory locus which controls exoenzyme S synthesis in Pseudomonas aeruginosa

T L Yahr and D W Frank

Journal of bacteriology, Vol.176(13), pp.3832-3838

07/1994

DOI: 10.1128/jb.176.13.3832-3838.1994

PMCID: PMC205579

PMID: 8021164

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Abstract

The transcriptional organization of the exoenzyme S trans-regulatory locus was studied by using promoter fusion and transcriptional start site mapping analyses. The 5' regions flanking open reading frames encoding ExsC, ExsB, ExsA, and ExsD were cloned in both orientations into the promoter vector pQF26, which contains the chloramphenicol acetyltransferase reporter gene (cat). CAT activity from each promoter fusion transformed into Pseudomonas aeruginosa and Escherichia coli was measured. The trans-regulatory locus promoters demonstrated low to undetectable CAT activity in E. coli regardless of the orientation of the DNA fragment relative to the reporter gene. In P. aeruginosa two of the promoter clones containing DNA located 5' of exsC (pC) and exsD (pD) demonstrated significant CAT activity. Transcriptional initiation from pC and pD was dependent on the orientation of the DNA fragment, the inclusion of a chelator in the growth medium, and the presence of a functional exsA gene. Transcriptional start sites were mapped for the pC and pD promoter regions by using total RNA isolated from P. aeruginosa strains grown in medium including a chelator. Our data are consistent with an operon model for the transcriptional organization of the exoenzyme S trans-regulatory locus. In addition, ExsA appears to be involved in controlling transcriptional initiation from both the trans-regulatory locus and a region located immediately downstream of the exsA gene.

Genes, Bacterial

RNA, Messenger - genetics

Transcriptional Activation

Molecular Sequence Data

Operon - genetics

Promoter Regions, Genetic - genetics

Recombinant Fusion Proteins

Gene Expression Regulation, Bacterial - drug effects

Nitrilotriacetic Acid - pharmacology

Poly(ADP-ribose) Polymerases - genetics

Pseudomonas aeruginosa - genetics

Signal Transduction - drug effects

Bacterial Toxins

Cations - pharmacology

Pseudomonas aeruginosa - enzymology

ADP Ribose Transferases

Base Sequence

Cloning, Molecular

Enzyme Induction - drug effects

Transcription, Genetic

Genes, Reporter

Details

Title: Subtitle: Transcriptional organization of the trans-regulatory locus which controls exoenzyme S synthesis in Pseudomonas aeruginosa
Creators: T L Yahr - Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226
D W Frank
Resource Type: Journal article
Publication Details: Journal of bacteriology, Vol.176(13), pp.3832-3838
DOI: 10.1128/jb.176.13.3832-3838.1994
PMID: 8021164
PMCID: PMC205579
NLM abbreviation: J Bacteriol
ISSN: 0021-9193
eISSN: 1098-5530
Publisher: United States
Grant note: R29-AI31665 / NIAID NIH HHS
Language: English
Date published: 07/1994
Academic Unit: Microbiology and Immunology
Record Identifier: 9984002376702771

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