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Journal of Bacteriology, July 2000, p. 3913-3919, Vol. 182, No. 14
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Regulatory RNA (PrrB RNA) Modulates Expression of Secondary Metabolite Genes in Pseudomonas fluorescens F113

Simon Aarons,dagger Abdelhamid Abbas, Claire Adams, Anne Fenton,Dagger and Fergal O'Gara*

BIOMERIT Research Centre, Department of Microbiology, National University of Ireland, Cork, Cork, Ireland

Received 10 January 2000/Accepted 24 April 2000

The GacS-GacA two-component signal transduction system, which is highly conserved in gram-negative bacteria, is required for the production of exoenzymes and secondary metabolites in Pseudomonas spp. Screening of a Pseudomonas fluorescens F113 gene bank led to the isolation of a previously undefined locus which could restore secondary metabolite production to both gacS and gacA mutants of F113. Sequence analysis of this locus demonstrated that it did not contain any obvious Pseudomonas protein-coding open reading frames or homologues within available databases. Northern analysis indicated that the locus encodes an RNA (PrrB RNA) which is able to phenotypically complement gacS and gacA mutants and is itself regulated by the GacS-GacA two-component signal transduction system. Primer extension analysis of the 132-base transcript identified the transcription start site located downstream of a sigma 70 promoter sequence from positions -10 to -35. Inactivation of the prrB gene in F113 resulted in a significant reduction of 2,4-diacetylphloroglucinol (Phl) and hydrogen cyanide (HCN) production, while increased metabolite production was observed when prrB was overexpressed. The prrB gene sequence contains a number of imperfect repeats of the consensus sequence 5'-AGGA-3', and sequence analysis predicted a complex secondary structure featuring multiple putative stem-loops with the consensus sequences predominantly positioned at the single-stranded regions at the ends of the stem-loops. This structure is similar to the CsrB and RsmB regulatory RNAs in Escherichia coli and Erwinia carotovora, respectively. Results suggest that a regulatory RNA molecule is involved in GacA-GacS-mediated regulation of Phl and HCN production in P. fluorescens F113.

* Corresponding author. Mailing address: BIOMERIT Research Centre, Department of Microbiology, National University of Ireland, Cork, Cork, Ireland. Phone: 353-21-272097. Fax: 353-21-275934. E-mail: f.ogara{at}bureau.ucc.ie.

dagger Present address: Cork Cancer Research Centre, Mercy Hospital, Cork, Ireland.

Dagger Present address: Department of Food Technology, National University of Ireland, Cork, Cork, Ireland.

Journal of Bacteriology, July 2000, p. 3913-3919, Vol. 182, No. 14
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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