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Journal of Bacteriology, November 2001, p. 6676-6683, Vol. 183, No. 22
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.22.6676-6683.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
The Global Posttranscriptional Regulator RsmA
Modulates Production of Virulence Determinants and
N-Acylhomoserine Lactones in Pseudomonas
aeruginosa
Gabriella
Pessi,1,
Faye
Williams,2,3
Zoë
Hindle,2,
Karin
Heurlier,1
Matthew T. G.
Holden,2,3,§
Miguel
Cámara,2
Dieter
Haas,1 and
Paul
Williams2,3,*
Laboratoire de Biologie Microbienne,
Université de Lausanne, CH-1015 Lausanne,
Switzerland,1 and School of
Pharmaceutical Sciences, University of Nottingham, University Park,
Nottingham NG7 2RD,2 and Institute
of Infections and Immunity, University of Nottingham, Queen's Medical
Centre, Nottingham NG7 2UH,3 United Kingdom
Received 12 April 2001/Accepted 20 August 2001
Posttranscriptional control is known to contribute to the
regulation of secondary metabolism and virulence determinants in certain gram-negative bacteria. Here we report the isolation of a
Pseudomonas aeruginosa gene which encodes a global
translational regulatory protein, RsmA (regulator of secondary
metabolites). Overexpression of rsmA resulted in a
substantial reduction in the levels of extracellular products,
including protease, elastase, and staphylolytic (LasA protease)
activity as well as the PA-IL lectin, hydrogen cyanide (HCN), and the
phenazine pigment pyocyanin. While inactivation of rsmA
in P. aeruginosa had only minor effects on the
extracellular enzymes and the PA-IL lectin, the production of HCN and
pyocyanin was enhanced during the exponential phase. The influence of
RsmA on N-acylhomoserine lactone-mediated quorum sensing
was determined by assaying the levels of
N-(3-oxododecanoyl)homoserine lactone (3-oxo-C12-HSL)
and N-butanoylhomoserine lactone (C4-HSL) produced by
the rsmA mutant and the
rsmA-overexpressing strain. RsmA exerted a negative
effect on the synthesis of both 3-oxo-C12-HSL and C4-HSL, which was
confirmed by using lasI and rhlI
translational fusions. These data also highlighted the temporal
expression control of the lasI gene, which was induced
much earlier and to a higher level during the exponential growth phase
in an rsmA mutant. To investigate whether RsmA modulates
HCN production solely via quorum-sensing control, hcn
translational fusions were employed to monitor the regulation of the
cyanide biosynthesis genes (hcnABC). RsmA was shown to
exert an additional negative effect on cyanogenesis
posttranscriptionally by acting on a region surrounding the
hcnA ribosome-binding site. This suggests that, in
P. aeruginosa, RsmA functions as a pleiotropic posttranscriptional regulator of secondary metabolites directly and
also indirectly by modulating the quorum-sensing circuitry.
*
Corresponding author. Mailing address: School of
Pharmaceutical Sciences, University of Nottingham, University Park,
Nottingham NG7 2RD, United Kingdom. Phone: 44 115 9515047. Fax: 44 115 8466296. E-mail: paul.williams{at}nottingham.ac.uk.

Present address: Nestlé Research Center, CH-1000 Lausanne
26,
Switzerland.

Present address: Microscience Ltd., Wokingham, Berks., RG41
5TU, United
Kingdom.
§
Present address: The Sanger Centre, Wellcome Trust Genome Campus,
Hinxton, Cambridge CB10 1SA, United
Kingdom.
Journal of Bacteriology, November 2001, p. 6676-6683, Vol. 183, No. 22
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.22.6676-6683.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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