Faculty Collaboration Database

Transcriptional organization of the trans-regulatory locus which controls exoenzyme S synthesis in Pseudomonas aeruginosa. J Bacteriol 1994 Jul;176(13):3832-38

Date

07/01/1994

Pubmed ID

8021164

Pubmed Central ID

PMC205579

DOI

10.1128/jb.176.13.3832-3838.1994

Scopus ID

2-s2.0-0028332195 (requires institutional sign-in at Scopus site)   76 Citations

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.

Author List

Yahr TL, Frank DW

Author

Dara W. Frank PhD Professor in the Microbiology and Immunology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

ADP Ribose Transferases
Bacterial Toxins
Base Sequence
Cations
Cloning, Molecular
Enzyme Induction
Gene Expression Regulation, Bacterial
Genes, Bacterial
Genes, Reporter
Molecular Sequence Data
Nitrilotriacetic Acid
Operon
Poly(ADP-ribose) Polymerases
Promoter Regions, Genetic
Pseudomonas aeruginosa
RNA, Messenger
Recombinant Fusion Proteins
Signal Transduction
Transcription, Genetic
Transcriptional Activation