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Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins

Horizontal gene transfer is a major driver of bacterial evolution and adaptation to environmental stresses, occurring notably via transformation of naturally competent organisms. The Deinococcus radiodurans bacterium, characterized by its extreme radioresistance, is also naturally competent. Here, w...

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Autores principales: Ithurbide, Solenne, Coste, Geneviève, Lisboa, Johnny, Eugénie, Nicolas, Bentchikou, Esma, Bouthier de la Tour, Claire, Liger, Dominique, Confalonieri, Fabrice, Sommer, Suzanne, Quevillon-Cheruel, Sophie, Servant, Pascale
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7314969/
https://www.ncbi.nlm.nih.gov/pubmed/32625182
http://dx.doi.org/10.3389/fmicb.2020.01253
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author Ithurbide, Solenne
Coste, Geneviève
Lisboa, Johnny
Eugénie, Nicolas
Bentchikou, Esma
Bouthier de la Tour, Claire
Liger, Dominique
Confalonieri, Fabrice
Sommer, Suzanne
Quevillon-Cheruel, Sophie
Servant, Pascale
author_facet Ithurbide, Solenne
Coste, Geneviève
Lisboa, Johnny
Eugénie, Nicolas
Bentchikou, Esma
Bouthier de la Tour, Claire
Liger, Dominique
Confalonieri, Fabrice
Sommer, Suzanne
Quevillon-Cheruel, Sophie
Servant, Pascale
author_sort Ithurbide, Solenne
collection PubMed
description Horizontal gene transfer is a major driver of bacterial evolution and adaptation to environmental stresses, occurring notably via transformation of naturally competent organisms. The Deinococcus radiodurans bacterium, characterized by its extreme radioresistance, is also naturally competent. Here, we investigated the role of D. radiodurans players involved in different steps of natural transformation. First, we identified the factors (PilQ, PilD, type IV pilins, PilB, PilT, ComEC-ComEA, and ComF) involved in DNA uptake and DNA translocation across the external and cytoplasmic membranes and showed that the DNA-uptake machinery is similar to that described in the Gram negative bacterium Vibrio cholerae. Then, we studied the involvement of recombination and DNA repair proteins, RecA, RecF, RecO, DprA, and DdrB into the DNA processing steps of D. radiodurans transformation by plasmid and genomic DNA. The transformation frequency of the cells devoid of DprA, a highly conserved protein among competent species, strongly decreased but was not completely abolished whereas it was completely abolished in ΔdprA ΔrecF, ΔdprA ΔrecO, and ΔdprA ΔddrB double mutants. We propose that RecF and RecO, belonging to the recombination mediator complex, and DdrB, a specific deinococcal DNA binding protein, can replace a function played by DprA, or alternatively, act at a different step of recombination with DprA. We also demonstrated that a ΔdprA mutant is as resistant as wild type to various doses of γ-irradiation, suggesting that DprA, and potentially transformation, do not play a major role in D. radiodurans radioresistance.
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spelling pubmed-73149692020-07-02 Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins Ithurbide, Solenne Coste, Geneviève Lisboa, Johnny Eugénie, Nicolas Bentchikou, Esma Bouthier de la Tour, Claire Liger, Dominique Confalonieri, Fabrice Sommer, Suzanne Quevillon-Cheruel, Sophie Servant, Pascale Front Microbiol Microbiology Horizontal gene transfer is a major driver of bacterial evolution and adaptation to environmental stresses, occurring notably via transformation of naturally competent organisms. The Deinococcus radiodurans bacterium, characterized by its extreme radioresistance, is also naturally competent. Here, we investigated the role of D. radiodurans players involved in different steps of natural transformation. First, we identified the factors (PilQ, PilD, type IV pilins, PilB, PilT, ComEC-ComEA, and ComF) involved in DNA uptake and DNA translocation across the external and cytoplasmic membranes and showed that the DNA-uptake machinery is similar to that described in the Gram negative bacterium Vibrio cholerae. Then, we studied the involvement of recombination and DNA repair proteins, RecA, RecF, RecO, DprA, and DdrB into the DNA processing steps of D. radiodurans transformation by plasmid and genomic DNA. The transformation frequency of the cells devoid of DprA, a highly conserved protein among competent species, strongly decreased but was not completely abolished whereas it was completely abolished in ΔdprA ΔrecF, ΔdprA ΔrecO, and ΔdprA ΔddrB double mutants. We propose that RecF and RecO, belonging to the recombination mediator complex, and DdrB, a specific deinococcal DNA binding protein, can replace a function played by DprA, or alternatively, act at a different step of recombination with DprA. We also demonstrated that a ΔdprA mutant is as resistant as wild type to various doses of γ-irradiation, suggesting that DprA, and potentially transformation, do not play a major role in D. radiodurans radioresistance. Frontiers Media S.A. 2020-06-18 /pmc/articles/PMC7314969/ /pubmed/32625182 http://dx.doi.org/10.3389/fmicb.2020.01253 Text en Copyright © 2020 Ithurbide, Coste, Lisboa, Eugénie, Bentchikou, Bouthier de la Tour, Liger, Confalonieri, Sommer, Quevillon-Cheruel and Servant. http://creativecommons.org/licenses/by/4.0/ 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.
spellingShingle Microbiology
Ithurbide, Solenne
Coste, Geneviève
Lisboa, Johnny
Eugénie, Nicolas
Bentchikou, Esma
Bouthier de la Tour, Claire
Liger, Dominique
Confalonieri, Fabrice
Sommer, Suzanne
Quevillon-Cheruel, Sophie
Servant, Pascale
Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins
title Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins
title_full Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins
title_fullStr Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins
title_full_unstemmed Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins
title_short Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins
title_sort natural transformation in deinococcus radiodurans: a genetic analysis reveals the major roles of dpra, ddrb, reca, recf, and reco proteins
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7314969/
https://www.ncbi.nlm.nih.gov/pubmed/32625182
http://dx.doi.org/10.3389/fmicb.2020.01253
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