Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus

© 2018 Agúndez et al.

Detalles Bibliográficos
Autores: Agúndez, Leticia, Zárate-Pérez, Francisco, Meier, Anita F., Bardelli, Martino, Llosa, Matxalen, Escalante, Carlos R., Linden, R. Michael, Henckaerts, Els
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2018
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/248999
Acceso en línea:http://hdl.handle.net/10261/248999
Access Level:acceso abierto
Palabra clave:DNA-binding proteins
Plasmid construction
Transfection
Protein domains
DNA helicases
Helicases
Sedimentation
DNA replication
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spelling Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virusAgúndez, LeticiaZárate-Pérez, FranciscoMeier, Anita F.Bardelli, MartinoLlosa, MatxalenEscalante, Carlos R.Linden, R. MichaelHenckaerts, ElsDNA-binding proteinsPlasmid constructionTransfectionProtein domainsDNA helicasesHelicasesSedimentationDNA replication© 2018 Agúndez et al.Endonucleases of the HUH family are specialized in processing single-stranded DNA in a variety of evolutionarily highly conserved biological processes related to mobile genetic elements. They share a structurally defined catalytic domain for site-specific nicking and strand-transfer reactions, which is often linked to the activities of additional functional domains, contributing to their overall versatility. To assess if these HUH domains could be interchanged, we created a chimeric protein from two distantly related HUH endonucleases, containing the N-terminal HUH domain of the bacterial conjugative relaxase TrwC and the C-terminal DNA helicase domain of the human adeno-associated virus (AAV) replicase and site-specific integrase. The purified chimeric protein retained oligomerization properties and DNA helicase activities similar to Rep68, while its DNA binding specificity and cleaving-joining activity at oriT was similar to TrwC. Interestingly, the chimeric protein could catalyse site-specific integration in bacteria with an efficiency comparable to that of TrwC, while the HUH domain of TrwC alone was unable to catalyze this reaction, implying that the Rep68 C-terminal helicase domain is complementing the TrwC HUH domain to achieve site-specific integration into TrwC targets in bacteria. Our results illustrate how HUH domains could have acquired through evolution other domains in order to attain new roles, contributing to the functional flexibility observed in this protein superfamily.This work was supported by the Medical Research Council (MRC) grant MR/N022890/1 to EH and grant 1001764 to RML; National Institutes of Health (NIH) grant RO1-GM09285 to CRE; Spanish Ministry of Economy and Competitiveness (MINECO) grant BIO2013-46414-P to ML and AFM is supported by a Doc.Mobility fellowship from the Swiss National Science Foundation.Public Library of ScienceConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2021202120182021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/248999reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1371/journal.pone.0200841Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2489992026-05-22T06:33:51Z
dc.title.none.fl_str_mv Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
title Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
spellingShingle Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
Agúndez, Leticia
DNA-binding proteins
Plasmid construction
Transfection
Protein domains
DNA helicases
Helicases
Sedimentation
DNA replication
title_short Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
title_full Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
title_fullStr Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
title_full_unstemmed Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
title_sort Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus
dc.creator.none.fl_str_mv Agúndez, Leticia
Zárate-Pérez, Francisco
Meier, Anita F.
Bardelli, Martino
Llosa, Matxalen
Escalante, Carlos R.
Linden, R. Michael
Henckaerts, Els
author Agúndez, Leticia
author_facet Agúndez, Leticia
Zárate-Pérez, Francisco
Meier, Anita F.
Bardelli, Martino
Llosa, Matxalen
Escalante, Carlos R.
Linden, R. Michael
Henckaerts, Els
author_role author
author2 Zárate-Pérez, Francisco
Meier, Anita F.
Bardelli, Martino
Llosa, Matxalen
Escalante, Carlos R.
Linden, R. Michael
Henckaerts, Els
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv DNA-binding proteins
Plasmid construction
Transfection
Protein domains
DNA helicases
Helicases
Sedimentation
DNA replication
topic DNA-binding proteins
Plasmid construction
Transfection
Protein domains
DNA helicases
Helicases
Sedimentation
DNA replication
description © 2018 Agúndez et al.
publishDate 2018
dc.date.none.fl_str_mv 2018
2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/248999
url http://hdl.handle.net/10261/248999
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1371/journal.pone.0200841
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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