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Artificial sRNAs activating the Gac/Rsm signal transduction pathway in Pseudomonas fluorescens

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Abstract

In Pseudomonas fluorescens CHA0, the synthesis of antifungal compounds is post-transcriptionally activated by the Gac/Rsm cascade. The two-component system GacS/GacA promotes transcription of three small regulatory RNAs (i.e., sRNAs), RsmX, RsmY, and RsmZ, which remove the regulatory proteins RsmA and RsmE from the ribosome-binding sites of exoproduct-related mRNAs. The GacS/GacA-dependent accumulation of RsmX/Y/Z and formation of RsmX/Y/Z-RsmA/E complexes relieve mRNA translational repression. Other bacteria as E. coli and Vibrio spp. utilize similar sRNA–protein based systems to adjust mRNA translation (e.g., the E. coli Csr system for carbon storage, motility and biofilm regulation). The Rsm/Csr sRNAs are remarkably similar in that they contain several stem-loops with an invariant GGA trinucleotide exposed in the hairpin loop that would be the characteristic structural-sequence motifs relevant for sRNA activity and stability. Here it is shown that the dysfunctional Gac/Rsm cascade of P. fluorescens ΔrsmXYZ mutants could be restored by appropriate transcription levels of artificial genes encoding RNAs with unrelated primary sequence but with two or more hairpins displaying the RsmA/E binding motifs. The results support the hypothesis that the molecular mimicry of Rsm/Csr sRNAs is based on proper secondary structures that expose critical binding motifs irrespective of their overall sequence.

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Abbreviations

Gac:

Global antibiotic and cyanide control

Rsm:

Regulator of secondary metabolism

sRNAs:

Small non-coding regulatory RNAs

arsm :

Artificial rsm-like RNA gene

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Acknowledgments

This work was supported by grants from Banco RIO, CONICET and Universidad Nacional de Quilmes (Argentina). The author is grateful to Prof. Dieter Haas (DMF, Université de Lausanne, Switzerland) for providing P. fluorescens strains. The author would like to acknowledge the critical comments of two anonymous reviewers which contributed to improve this manuscript. C. V. is member of CONICET (Argentina).

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Authors and Affiliations

  1. Programa Interacciones Biológicas, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal, B1876BXD, Buenos Aires, Argentina

    Claudio Valverde

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  1. Claudio Valverde
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Correspondence to Claudio Valverde.

Additional information

Communicated by John Helmann.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Figure S1:

Stability of RsmY-like RNA homologues expressed in P. fluorescens. (JPG 183 kb)

Supplementary Figure S2:

Biological activity of RsmY-like RNA homologues. (JPG 293 kb)

Supplementary file 1:

Sequence of studied RNA molecules and comparative analysis of CsrA/RsmA recognition motifs. (DOC 76 kb)

Supplementary file 2:

Nucleotide substitutions in hairpins of RsmZ RNA tend to preserve secondary structure. (DOC 220 kb)

Supplementary Tables (PDF 29 kb)

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Valverde, C. Artificial sRNAs activating the Gac/Rsm signal transduction pathway in Pseudomonas fluorescens . Arch Microbiol 191, 349–359 (2009). https://doi.org/10.1007/s00203-009-0459-x

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