Identification of small RNAs associated with RNA chaperone Hfq reveals a new stress response regulator in Actinobacillus pleuropneumoniae

Giarlã Cunha da Silva, Ciro César Rossi, Jéssica Nogueira Rosa, Newton Moreno Sanches, Daniela Lopes Cardoso, Yanwen Li, Adam A. Witney, Kate A. Gould, Patrícia Pereira Fontes, Anastasia J. Callaghan, Janine Thérèse Bossé, Paul Richard Langford, Denise Mara Soares Bazzolli

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Abstract

The RNA chaperone Hfq promotes the association of small RNAs (sRNAs) with cognate mRNAs, controlling the expression of bacterial phenotype. Actinobacillus pleuropneumoniae hfq mutants strains are attenuated for virulence in pigs, impaired in the ability to form biofilms, and more susceptible to stress, but knowledge of the extent of sRNA involvement is limited. Here, using A. pleuropneumoniae strain MIDG2331 (serovar 8), 14 sRNAs were identified by co-immunoprecipitation with Hfq and the expression of eight, identified as trans-acting sRNAs, were confirmed by Northern blotting. We focused on one of these sRNAs, named Rna01, containing a putative promoter for RpoE (stress regulon) recognition. Knockout mutants of rna01 and a double knockout mutant of rna01 and hfq, both had decreased biofilm formation and hemolytic activity, attenuation for virulence in Galleria mellonella, altered stress susceptibility, and an altered outer membrane protein profile. Rna01 affected extracellular vesicle production, size and toxicity in G. mellonella. qRT-PCR analysis of rna01 and putative cognate mRNA targets indicated that Rna01 is associated with the extracytoplasmic stress response. This work increases our understanding of the multilayered and complex nature of the influence of Hfq-dependent sRNAs on the physiology and virulence of A. pleuropneumoniae.
Original languageEnglish
Article number1017278
Number of pages20
JournalFrontiers in Microbiology
Volume13
DOIs
Publication statusPublished - 4 Oct 2022

Keywords

  • Pasteurellaceae
  • porcine pleuropneumonia
  • trans-acting small RNA
  • extracellular vesicles
  • Galleria mellonella
  • UKRI
  • BBSRC
  • BB/K021109/1
  • BB/G019177/1
  • BB/M023052/1
  • BB/M020576/1
  • BB/S020543/1
  • BB/P001262/1
  • BB/G018553/1

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