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J. Biol. Chem., Vol. 277, Issue 1, 462-468, January 4, 2002
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From the Rhizobium etli CNPAF512 produces an
autoinducer that inhibits growth of Rhizobium
leguminosarum bv. viciae 248 and activates the
Agrobacterium tumefaciens tra reporter system. Production of this compound in R. etli is dependent on two genes,
named cinR and cinI, postulated to code for a
transcriptional regulator and an autoinducer synthase, respectively.
NMR analysis of the purified molecule indicates that the R. etli autoinducer produced by CinI is a saturated long chain
3-hydroxy-acyl-homoserine lactone, abbreviated as 3OH-(slc)-HSL. Using
cin-gusA fusions, expression of
cinI and cinR was shown to be growth
phase-dependent. Deletion analysis of the cinI
promoter region indicates that a regulatory element negatively controls
cinI expression. Mutational analysis revealed that
expression of the cinI gene is positively regulated by the CinR/3OH-(slc)-HSL complex. Besides 3OH-(slc)-HSL, R. etli
produces at least six other autoinducer molecules, for which the
structures have not yet been revealed, and of which the synthesis
requires the previously identified raiI and
raiR genes. At least three different autoinducers,
including a compound co-migrating with 3OH-(slc)-HSL, are produced in
R. etli bacteroids isolated from bean nodules. This is
further substantiated by the observation that cinI and
cinR are both expressed under symbiotic conditions. Acetylene reduction activity of nodules induced by the cin
mutants was reduced with 60-70% compared with wild-type nodules,
indicating that the R. etli 3OH-(slc)-HSL is involved in
the symbiotic process. This was further confirmed by transmission
electron microscopy of nodules induced by the wild type and the
cinI mutant. Symbiosomes carrying cinI mutant
bacteroids did not fully differentiate compared with wild-type
symbiosomes. Finally, it was observed that the cinR
gene and raiR control growth of R. etli.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF393621.
The cin Quorum Sensing Locus of Rhizobium
etli CNPAF512 Affects Growth and Symbiotic Nitrogen Fixation*
§,,,,,,
,**, and
Centre of Microbial and Plant Genetics,
Katholicke Universitat Leuven, Kasteelpark Arenberg 20, B-3001
Heverlee, Belgium, the ¶ Centre for Surface Chemistry and
Catalysis, Katholicke Universitat Leuven, Kasteelpark Arenberg 23, B-3001 Heverlee, Belgium, and the Zoological Institute,
Katholicke Universitat Leuven, Naamsestraat 59, B-3000 Leuven,
Belgium
*
Parts of this study were presented at the American
Society for Microbiology Conference on Cell-Cell Communication in
Bacteria, July 6-9, 2001, Snowbird, UT.The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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