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

Ithurbide, S; Coste, G; Lisboa, J; Eugénie, N; Bentchikou, E; Bouthier de la Tour, C; Liger, D; Confalonieri, F; Sommer, S; Quevillon-Cheruel, S; Servant, P

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

Coste, G Lisboa, J Eugénie, N Bentchikou, E Bouthier de la Tour, C Liger, D Confalonieri, F Sommer, S Quevillon-Cheruel, S Servant, P

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Publisher:: FRONTIERS MEDIA SA
Publication Type:: Journal Article
Citation:: Frontiers in microbiology, 2020, 11, pp. 1253
Issue Date:: 2020-01

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Field	Value	Language
dc.contributor.author	Ithurbide, S https://orcid.org/0000-0002-1533-8472
dc.contributor.author	Coste, G
dc.contributor.author	Lisboa, J
dc.contributor.author	Eugénie, N
dc.contributor.author	Bentchikou, E
dc.contributor.author	Bouthier de la Tour, C
dc.contributor.author	Liger, D
dc.contributor.author	Confalonieri, F
dc.contributor.author	Sommer, S
dc.contributor.author	Quevillon-Cheruel, S
dc.contributor.author	Servant, P
dc.date.accessioned	2020-08-17T07:01:49Z
dc.date.available	2020-05-18
dc.date.available	2020-08-17T07:01:49Z
dc.date.issued	2020-01
dc.identifier.citation	Frontiers in microbiology, 2020, 11, pp. 1253
dc.identifier.issn	1664-302X
dc.identifier.issn	1664-302X
dc.identifier.uri	http://hdl.handle.net/10453/142159
dc.description.abstract	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.
dc.format	Electronic-eCollection
dc.language	eng
dc.publisher	FRONTIERS MEDIA SA
dc.relation.ispartof	Frontiers in microbiology
dc.relation.isbasedon	10.3389/fmicb.2020.01253
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	0502 Environmental Science and Management, 0503 Soil Sciences, 0605 Microbiology
dc.title	Natural Transformation in Deinococcus radiodurans: A Genetic Analysis Reveals the Major Roles of DprA, DdrB, RecA, RecF, and RecO Proteins.
dc.type	Journal Article
utslib.citation.volume	11
utslib.location.activity	Switzerland
utslib.for	0502 Environmental Science and Management
utslib.for	0503 Soil Sciences
utslib.for	0605 Microbiology
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
pubs.organisational-group	/University of Technology Sydney
utslib.copyright.status	open_access	*
dc.date.updated	2020-08-17T07:01:28Z
pubs.publication-status	Published
pubs.volume	11

Abstract:

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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http://hdl.handle.net/10453/142159