K. Winzer et al., The Pseudomonas aeruginosa lectins PA-IL and PA-IIL are controlled by quorum sensing and by RpoS, J BACT, 182(22), 2000, pp. 6401-6411
In Pseudomonas aeruginosa, many exoproduct virulence determinants are regul
ated via a hierarchical quorum-sensing cascade involving the transcriptiona
l regulators LasR and RhlR and their cognate activators, N-(3-oxododecanoyl
)-L-homoserine lactone (30-C12-HSL) and N-butanoyl-L-homoserine lactone (C4
-HSL). In this paper, we demonstrate that the cytotoxic lectins PA-IL and P
A-IIL are regulated via quorum sensing. Using immunoblot analysis, the prod
uction of both lectins was found to be directly dependent on the rhl locus
while, in a lasR mutant, the onset of lectin synthesis was delayed but not
abolished. The PA-IL structural gene, lecA, was cloned and sequenced. Trans
cript analysis indicated a monocistronic organization with a transcriptiona
l start site 70 bp upstream of the lecA translational start codon. A lux bo
x-type element together with RpoS (sigma (S)) consensus sequences was ident
ified upstream of the putative promoter region. In Escherichia coli, expres
sion of a lecA::lux reporter fusion was activated by RhlR/C4-HSL, but not b
y LasR/3O-C12-HSL, confirming direct regulation by RhlR/C4-HSL. Similarly,
in P. aeruginosa PAO1, the expression of a chromosomal lecA::lux fusion was
enhanced but not advanced by the addition of exogenous C4-HSL but not 30-C
12-HSL. Furthermore, mutation of rpoS abolished lectin synthesis in P. aeru
ginosa, demonstrating that both RpoS and RhlR/C4-HSL are required. Although
the C4-HSL-dependent expression of the lecA::lux reporter in E. coli could
be inhibited by the presence of 30-C12-HSL, this did not occur in P. aerug
inosa. This suggests that, in the homologous genetic background, 30-C12-HSL
does not function as a posttranslational regulator of the RhlR/C4-HSL-depe
ndent activation of lecA expression.