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A Single Mutation in the oprF mRNA Leader Confers Strict Translational Control by the Gac/Rsm System in Pseudomonas fluorescens CHA0

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Abstract

The rhizobacterium Pseudomonas fluorescens CHA0 is able to antagonize fungal phytopathogens on a variety of crop plants, mainly due to the production of secondary metabolites that are coordinately upregulated by the global regulatory Gac/Rsm cascade. The two-component system GacS/GacA activates transcription of the three small regulatory RNAs RsmX, RsmY, and RsmZ, which counteract translational repression of target mRNAs by RsmA and RsmE proteins. In a search for novel Gac/Rsm targets based on the minimal sequence on mRNA leaders required for RsmA/RsmE control, the leader region of the major porin OprF emerged as a candidate. Although an isogenic CHA0 oprF mutant showed a reduced ability to attach to cucumber and tomato roots, suggesting a role for OprF in root colonization as a requisite for pathogen antagonism, a translational oprF′-′lacZ fusion was weakly regulated by Gac/Rsm despite its high sequence similarity to the hcnA leader. A single base substitution put the modified oprF 5′-UTR under strict control by Gac/Rsm. The results highlight the subtle sequence requirements of Gac/Rsm targets.

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Authors and Affiliations

  1. Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal, B1876BXD, Argentina

    María Cecilia Alvarez Crespo & Claudio Valverde

Authors
  1. María Cecilia Alvarez Crespo
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  2. Claudio Valverde
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Correspondence to Claudio Valverde.

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Supplementary Table 1

MOESM1 Bacterial strains, plasmids and oligonucleotides used in this study (DOC 70 kb)

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Alvarez Crespo, M.C., Valverde, C. A Single Mutation in the oprF mRNA Leader Confers Strict Translational Control by the Gac/Rsm System in Pseudomonas fluorescens CHA0. Curr Microbiol 58, 182–188 (2009). https://doi.org/10.1007/s00284-008-9306-6

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  • DOI: https://doi.org/10.1007/s00284-008-9306-6

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