Document Detail

Triggering the ExoS regulon of Pseudomonas aeruginosa: A GFP-reporter analysis of exoenzyme (Exo) S, ExoT and ExoU synthesis.
MedLine Citation:
PMID:  11095918     Owner:  NLM     Status:  MEDLINE    
The ExoS regulon of Pseudomonas aeruginosa encodes diverse type III secreted effector proteins which have been shown to exert cytotoxic effects in cell culture experiments. However, little information exists about the environmental conditions and stimuli for upregulation of the ExoS regulon. Translational reporter fusion proteins of exoenzyme (Exo) S, ExoT and ExoU, as well as the type II secreted exotoxin A (ETA) to the green fluorescent protein (GFP), were constructed in order to compare exoprotein production under diverse growth conditions. Reporter protein activity was recorded by FACS-analysis and by conventional and confocal laser scanning microscopy. Low ion concentration induced co-ordinated upregulation of ExoS, ExoT and ExoU with a maximum effect at 37 degrees C. A dose-dependent upregulation was seen with human serum or increasing NaCl concentrations. A type III secretion-negative pcrD mutant of P. aeruginosa showed a weak ExoS response to environmental stimuli, compared with the parental strain, suggesting a negative regulatory mechanism. Co-culture with the mammalian cell lines J774A.1 or HeLa led to rapid upregulation of ExoS, ExoT and ExoU synthesis. These data suggest that the ExoS regulon of P. aeruginosa can be triggered by a variety of environmental signals as well as by cell contact with eukaryotic cells.
M W Hornef; A Roggenkamp; A M Geiger; M Hogardt; C A Jacobi; J Heesemann
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Microbial pathogenesis     Volume:  29     ISSN:  0882-4010     ISO Abbreviation:  Microb. Pathog.     Publication Date:  2000 Dec 
Date Detail:
Created Date:  2001-01-10     Completed Date:  2001-03-29     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  8606191     Medline TA:  Microb Pathog     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  329-43     Citation Subset:  IM    
Copyright Information:
Copyright 2000 Academic Press.
Max von Pettenkofer Institut, Ludwig Maximilian-Universität, Munich, Germany.
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MeSH Terms
ADP Ribose Transferases*
Bacterial Proteins / biosynthesis,  genetics
Bacterial Toxins*
Blotting, Southern
Chemiluminescent Measurements
Coculture Techniques
Conjugation, Genetic / physiology
DNA Primers / chemistry
Electrophoresis, Polyacrylamide Gel
Exotoxins / biosynthesis,  genetics
Flow Cytometry
Green Fluorescent Proteins
Hela Cells
Indicators and Reagents / chemistry
Luminescent Proteins / chemistry
Microscopy, Confocal
Microscopy, Fluorescence
Plasmids / chemistry
Polymerase Chain Reaction
Protein Kinases / biosynthesis*,  chemistry,  genetics
Pseudomonas aeruginosa / chemistry,  genetics,  pathogenicity*
Regulon / physiology*
Signal Transduction
Virulence Factors*
Reg. No./Substance:
0/Bacterial Proteins; 0/Bacterial Toxins; 0/DNA Primers; 0/Exotoxins; 0/Indicators and Reagents; 0/Luminescent Proteins; 0/Virulence Factors; 0/pseudomonas exoprotein A protein, Pseudomonas aeruginosa; 147336-22-9/Green Fluorescent Proteins; EC 2.4.2.-/ADP Ribose Transferases; EC protein, Pseudomonas aeruginosa; EC 2.7.-/Protein Kinases; EC 2.7.3.-/protein-histidine kinase

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