title: Functional analysis of the Pseudomonas aeruginosa autoinducer PAI.
abstract: A series of structural analogs of the Pseudomonas aeruginosa autoinducer [PAI, N-3-oxo-dodecanoyl homoserine lactone] were obtained and tested for their ability to act as autoinducers in stimulating the expression of the gene for elastase (lasB) by measuring beta-galactosidase production from a lasB-lacZ gene fusion in the presence of the transcriptional activator LasR. The data suggest that the length of the acyl side chain of the autoinducer molecule is the most critical factor for activity. Replacement of the ring O by S in the homoserine lactone moiety can be tolerated. Tritium-labelled PAI ([3H]PAI) was synthesized and used to demonstrate the association of [3H]PAI with cells overexpressing LasR. The PAI analogs were also tested for their ability to compete with [3H]PAI for binding of LasR. Results from the competition assays suggest that once again the length of the acyl side chain appears to be crucial for antagonist activity. The presence of the 3-oxo moiety also plays a significant role in binding since analogs which lacked this moiety were much less effective in blocking binding of [3H]PAI. All analogs demonstrating competition with PAI in binding to LasR also exhibited the ability to activate lasB expression, suggesting that they are functional analogs of PAI.
authors.affiliation: Department of Microbiology, University of Rochester, New York 14642, USA.
authors.forename: L
authors.initials: L
authors.lastname: Passador
authors.forename: K D
authors.initials: KD
authors.lastname: Tucker
authors.forename: K R
authors.initials: KR
authors.lastname: Guertin
authors.forename: M P
authors.initials: MP
authors.lastname: Journet
authors.forename: A S
authors.initials: AS
authors.lastname: Kende
authors.forename: B H
authors.initials: BH
authors.lastname: Iglewski
mesh_terms: D001426:Bacterial Proteins; D001667:Binding, Competitive; D004268:DNA-Binding Proteins / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006714:Homoserine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D007783:Lactones / Q000737:chemistry* / Q000378:metabolism; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D013329:Structure-Activity Relationship; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D007783:Lactones; C080365:LasR protein, Pseudomonas aeruginosa; D015534:Trans-Activators; C085113:Pseudomonas aeruginosa autoinducer; D006714:Homoserine; D008666:Metalloendopeptidases; C043859:pseudolysin, Pseudomonas aeruginosa