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

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Title: Identification of small RNAs associated with RNA chaperone Hfq reveals a new stress response regulator in Actinobacillus pleuropneumoniae
Authors: Cunha da Silva, G
Rossi, CC
Nogueira Rosa, J
Moreno Sanches, N
Lopes Cardoso, D
Li, Y
Witney, A
Gould, K
Pereira Fontes, P
Callaghan, A
Bosse, J
Langford, P
Bazzolli, D
Item Type: Journal Article
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.
Issue Date: 4-Oct-2022
Date of Acceptance: 9-Sep-2022
URI: http://hdl.handle.net/10044/1/99712
DOI: 10.3389/fmicb.2022.1017278
ISSN: 1664-302X
Publisher: Frontiers Media
Journal / Book Title: Frontiers in Microbiology
Volume: 13
Copyright Statement: © 2022 da Silva, Rossi, Rosa, Sanches, Cardoso, Li, Witney, Gould, Fontes, Callaghan, Bossé, Langford and Bazzolli. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Biotechnology and Biological Sciences Research Council
Biotechnology and Biological Sciences Research Council (BBSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
BBSRC
Biotechnology and Biological Sciences Research Council (BBSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: BB/G018553/1
BB/K021109/1
BB/G018553/1
BB/S020543/1
BB/M023052/1
BB/M023052/1
BB/S002103/1
BB/S005897/1
Keywords: 0502 Environmental Science and Management
0503 Soil Sciences
0605 Microbiology
Publication Status: Published
Article Number: 1017278
Online Publication Date: 2022-10-04
Appears in Collections:Department of Infectious Diseases
Faculty of Medicine



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