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PMID: 1902216 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Cloning and characterization of the Pseudomonas aeruginosa lasR gene, a transcriptional activator of elastase expression.

Journal of bacteriology ·Vol. 173 ·No. 9 ·1991-05-00 ·Pages 3000-9

Gambello MJ, Iglewski BH

Abstract

We report the discovery of the lasR gene, which positively regulates elastase expression in Pseudomonas aeruginosa PAO1. The lasR gene was cloned by its ability to restore a positive elastase phenotype in strain PA103, a strain which possesses the elastase structural gene (lasB) but fails to synthesize the enzyme. Nucleotide sequence analysis revealed an open reading frame of 716 nucleotides encoding a protein of approximately 27 kDa. A labeled LasR protein of 27 kDa was detected in Escherichia coli by using a T7 RNA polymerase expression system. A chromosomal deletion mutant of the lasR gene was constructed in PAO1 by gene replacement. This mutant (PAO-R1) is devoid of elastolytic activity and elastase antigen. The deduced amino acid sequence of LasR is 27% homologous to the positive activator LuxR of Vibrio fischeri and the suspected activator 28K-UvrC of E. coli. Northern (RNA) analysis of total cellular RNA from PAO1, PAO-R1, and PAO-R1 containing the lasR gene on a multicopy plasmid (pMG1.7) revealed that a functional lasR gene is required for transcription of the elastase structural gene (lasB).

Related Genes
lasR
MeSH Terms
Amino Acid Sequence Base Sequence Blotting, Northern Blotting, Southern Cloning, Molecular Escherichia coli/genetics,metabolism Gene Expression Regulation, Bacterial Genes, Bacterial/genetics Genes, Regulator/genetics Molecular Sequence Data Pancreatic Elastase/biosynthesis Pseudomonas aeruginosa/genetics Sequence Homology, Nucleic Acid Trans-Activators/chemistry Transcription, Genetic
Chemicals
Trans-Activators Pancreatic Elastase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gambello M J
Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, New York 14642.
Iglewski B H
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.173.9.3000-3009.1991
Published
1991-05-00
Pages
3000-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC207884
Subset
IM
Grants
PHS HHS · A125669 · United States
NIGMS NIH HHS · T32GM07356 · United States
Databases
GENBANK
M55263, M55488, M55489, M55490, M55491, M55492, M55493, M55494, M59425, M93064
External Links
PubMed source DOI: 10.1128/jb.173.9.3000-3009.1991 PMC Full Text View journal details
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