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Journal of Bacteriology, October 1998, p. 5398-5405, Vol. 180, No. 20
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Analogs of the Autoinducer 3-Oxooctanoyl-Homoserine Lactone
Strongly Inhibit Activity of the TraR Protein of
Agrobacterium tumefaciens
Jun
Zhu,1
John W.
Beaber,1
Margret I.
Moré,1
Clay
Fuqua,2
Anatol
Eberhard,3 and
Stephen
C.
Winans1,*
Section of Microbiology, Cornell University, Ithaca, New
York 14853,1
Department of Biology,
Trinity University, San Antonio, Texas
78212,2 and
Department of Chemistry,
Ithaca College, Ithaca, New York
148503
Received 23 March 1998/Accepted 28 July 1998
The TraR and TraI proteins of Agrobacterium tumefaciens
mediate cell-density-dependent expression of the Ti plasmid
tra regulon. TraI synthesizes the autoinducer pheromone
N-(3-oxooctanoyl)-L-homoserine lactone
(3-oxo-C8-HSL), while TraR is an
3-oxo-C8-HSL-responsive transcriptional activator. We have
compared the abilities of 3-oxo-C8-HSL and 32 related
compounds to activate expression of a TraR-regulated promoter. In a
strain that expresses wild-type levels of TraR, only
3-oxo-C8-HSL was strongly stimulatory, four compounds were detectably active only at high concentrations, and the remaining 28 compounds were inactive. Furthermore, many of these compounds were
potent antagonists. In contrast, almost all of these compounds were
stimulatory in a congenic strain that overexpresses TraR and no
compound was a potent antagonist. We propose a model in which
autoinducers enhance the affinity of TraR either for other TraR
monomers or for DNA binding sites and that overexpression of TraR
potentiates this interaction by mass action. Wild-type A. tumefaciens released a rather broad spectrum of autoinducers, including several that antagonize induction of a wild-type strain. However, under all conditions tested, 3-oxo-C8-HSL was more
abundant than any other analog, indicating that other released
autoinducers do not interfere with tra gene induction. We
conclude that (i) in wild-type strains, only 3-oxo-C8-HSL
significantly stimulates tra gene expression, while many
autoinducer analogs are potent antagonists; (ii) TraR overexpression
increases agonistic activity of autoinducer analogs, allowing sensitive
biodetection of many autoinducers; and (iii) autoinducer stimulatory
activity is potentiated by TraR overproduction, suggesting that
autoinducers may shift an equilibrium between TraR
monomers and dimers or oligomers. When autoinducer
specificities of other quorum-sensing proteins are tested, care should
be taken not to overexpress those proteins.
*
Corresponding author. Mailing address: Section of
Microbiology, Cornell University, Ithaca, NY 14853. Phone: (607)
255-2413. Fax: (607) 255-3904. E-mail: scw2{at}cornell.edu.
Journal of Bacteriology, October 1998, p. 5398-5405, Vol. 180, No. 20
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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