Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana

[EN] Improved bioinformatics tools for annotating gene function are becoming increasingly available, but such information must be considered theoretical until further experimental evidence proves it. In the work reported here, the genes for the main components of the translocons of the outer membran...

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Detalhes bibliográficos
Autores: Sáiz-Bonilla, María, Martín Merchán, Andrea, Pallás Benet, Vicente|||0000-0003-4954-989X, NAVARRO-BOHIGUES, JOSE-ANTONIO
Tipo de documento: artigo
Data de publicação:2022
País:España
Recursos:Universitat Politècnica de València (UPV)
Repositório:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Idioma:inglês
OAI Identifier:oai:riunet.upv.es:10251/193100
Acesso em linha:https://riunet.upv.es/handle/10251/193100
Access Level:Acceso aberto
Palavra-chave:Nicotiana benthamiana
Chloroplasts
Mitochondria
Translocon receptor
Toc
Tom
Protein transport
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spelling Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamianaSáiz-Bonilla, MaríaMartín Merchán, AndreaPallás Benet, Vicente|||0000-0003-4954-989XNAVARRO-BOHIGUES, JOSE-ANTONIONicotiana benthamianaChloroplastsMitochondriaTranslocon receptorTocTomProtein transport[EN] Improved bioinformatics tools for annotating gene function are becoming increasingly available, but such information must be considered theoretical until further experimental evidence proves it. In the work reported here, the genes for the main components of the translocons of the outer membrane of chloroplasts (Toc) and mitochondria (Tom), including preprotein receptors and protein-conducting channels of N. benthamiana, were identified. Sequence identity searches and phylogenetic relationships with functionally annotated sequences such as those of A. thaliana revealed that N. benthamiana orthologs mainly exist as recently duplicated loci. Only a Toc34 ortholog was found (NbToc34), while Toc159 receptor family was composed of four orthologs but somewhat different from those of A. thaliana. Except for NbToc90, the rest (NbToc120, NbToc159A and NbToc159B) had a molecular weight of about 150 kDa and an acidic domain similar in length. Only two orthologs of the Tom20 receptors, NbTom20-1 and NbTom20-2, were found. The number of the Toc and Tom receptor isoforms in N. benthamiana was comparable to that previously reported in tomato and what we found in BLAST searches in other species in the genera Nicotiana and Solanum. After cloning, the subcellular localization of N. benthamiana orthologs was studied, resulting to be identical to that of A. thaliana receptors. Phenotype analysis after silencing together with relative expression analysis in roots, stems and leaves revealed that, except for the Toc and Tom channel- forming components (NbToc75 and NbTom40) and NbToc34, functional redundancy could be observed either among Toc159 or mitochondrial receptors. Finally, heterodimer formation between NbToc34 and the NbToc159 family receptors was confirmed by two alternative techniques indicating that different Toc complexes could be assembled. Additional work needs to be addressed to know if this results in a functional specialization of each Toc complex.Funding This work was funded by grant PID2020-115571RB-I00 from the Spanish Agencia Estatal de Investigacion (AEI) and Fondo Europeo de Desarrollo Regional (FEDER). JN and MS-B are the recipients of a postdoctoral contract and a PhD fellowship from the Ministerio de Ciencia, Innovacion y Universidades of Spain, repectively.Frontiers Media SAInstituto Universitario Mixto de Biología Molecular y Celular de PlantasAgencia Estatal de InvestigaciónEuropean Regional Development FundMinisterio de Ciencia, Innovación y UniversidadesRepositorio Institucional de la Universitat Politècnica de València Riunet20222022-10-26journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://riunet.upv.es/handle/10251/193100reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)InglésengAgencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020 PID2020-115571RB-I00 LA MODIFICACION N6-METILADENOSINA (M6A) DEL RNA Y EL DIRECCIONAMIENTO DUAL A CLOROPLASTOS Y MITOCONDRIAS COMO MECANISMOS REGULADORES EN LA BIOLOGIA DE LOS VIRUS RNA DE PLANTASopen accesshttp://purl.org/coar/access_right/c_abf2Reconocimiento (by)http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/1931002026-06-13T07:49:27Z
dc.title.none.fl_str_mv Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
title Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
spellingShingle Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
Sáiz-Bonilla, María
Nicotiana benthamiana
Chloroplasts
Mitochondria
Translocon receptor
Toc
Tom
Protein transport
title_short Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
title_full Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
title_fullStr Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
title_full_unstemmed Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
title_sort Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana
dc.creator.none.fl_str_mv Sáiz-Bonilla, María
Martín Merchán, Andrea
Pallás Benet, Vicente|||0000-0003-4954-989X
NAVARRO-BOHIGUES, JOSE-ANTONIO
author Sáiz-Bonilla, María
author_facet Sáiz-Bonilla, María
Martín Merchán, Andrea
Pallás Benet, Vicente|||0000-0003-4954-989X
NAVARRO-BOHIGUES, JOSE-ANTONIO
author_role author
author2 Martín Merchán, Andrea
Pallás Benet, Vicente|||0000-0003-4954-989X
NAVARRO-BOHIGUES, JOSE-ANTONIO
author2_role author
author
author
dc.contributor.none.fl_str_mv Instituto Universitario Mixto de Biología Molecular y Celular de Plantas
Agencia Estatal de Investigación
European Regional Development Fund
Ministerio de Ciencia, Innovación y Universidades
Repositorio Institucional de la Universitat Politècnica de València Riunet
dc.subject.none.fl_str_mv Nicotiana benthamiana
Chloroplasts
Mitochondria
Translocon receptor
Toc
Tom
Protein transport
topic Nicotiana benthamiana
Chloroplasts
Mitochondria
Translocon receptor
Toc
Tom
Protein transport
description [EN] Improved bioinformatics tools for annotating gene function are becoming increasingly available, but such information must be considered theoretical until further experimental evidence proves it. In the work reported here, the genes for the main components of the translocons of the outer membrane of chloroplasts (Toc) and mitochondria (Tom), including preprotein receptors and protein-conducting channels of N. benthamiana, were identified. Sequence identity searches and phylogenetic relationships with functionally annotated sequences such as those of A. thaliana revealed that N. benthamiana orthologs mainly exist as recently duplicated loci. Only a Toc34 ortholog was found (NbToc34), while Toc159 receptor family was composed of four orthologs but somewhat different from those of A. thaliana. Except for NbToc90, the rest (NbToc120, NbToc159A and NbToc159B) had a molecular weight of about 150 kDa and an acidic domain similar in length. Only two orthologs of the Tom20 receptors, NbTom20-1 and NbTom20-2, were found. The number of the Toc and Tom receptor isoforms in N. benthamiana was comparable to that previously reported in tomato and what we found in BLAST searches in other species in the genera Nicotiana and Solanum. After cloning, the subcellular localization of N. benthamiana orthologs was studied, resulting to be identical to that of A. thaliana receptors. Phenotype analysis after silencing together with relative expression analysis in roots, stems and leaves revealed that, except for the Toc and Tom channel- forming components (NbToc75 and NbTom40) and NbToc34, functional redundancy could be observed either among Toc159 or mitochondrial receptors. Finally, heterodimer formation between NbToc34 and the NbToc159 family receptors was confirmed by two alternative techniques indicating that different Toc complexes could be assembled. Additional work needs to be addressed to know if this results in a functional specialization of each Toc complex.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022-10-26
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://riunet.upv.es/handle/10251/193100
url https://riunet.upv.es/handle/10251/193100
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Agencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020 PID2020-115571RB-I00 LA MODIFICACION N6-METILADENOSINA (M6A) DEL RNA Y EL DIRECCIONAMIENTO DUAL A CLOROPLASTOS Y MITOCONDRIAS COMO MECANISMOS REGULADORES EN LA BIOLOGIA DE LOS VIRUS RNA DE PLANTAS
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media SA
publisher.none.fl_str_mv Frontiers Media SA
dc.source.none.fl_str_mv reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
instname:Universitat Politècnica de València (UPV)
instname_str Universitat Politècnica de València (UPV)
reponame_str RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
collection RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,301603

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