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

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...

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Autores: Saiz-Bonilla, María, Martín Merchán, Andrea, Pallás Benet, Vicente, Navarro, José A.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/295536
Acceso en línea:http://hdl.handle.net/10261/295536
Access Level:acceso abierto
Palabra clave:Nicotiana benthamiana
Chloroplasts
Mitochondria
Translocon receptor
Toc
Tom
Protein transport
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oai_identifier_str oai:digital.csic.es:10261/295536
network_acronym_str ES
network_name_str España
repository_id_str
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
Saiz-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 Saiz-Bonilla, María
Martín Merchán, Andrea
Pallás Benet, Vicente
Navarro, José A.
author Saiz-Bonilla, María
author_facet Saiz-Bonilla, María
Martín Merchán, Andrea
Pallás Benet, Vicente
Navarro, José A.
author_role author
author2 Martín Merchán, Andrea
Pallás Benet, Vicente
Navarro, José A.
author2_role author
author
author
dc.contributor.none.fl_str_mv Agencia Estatal de Investigación (España)
European Commission
Ministerio de Ciencia, Innovación y Universidades (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
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 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
2023
2023
2023
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/10261/295536
url http://hdl.handle.net/10261/295536
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-115571RB-I00
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_4_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s012
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_3_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s011
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_2_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s010
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_1_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s009
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_7_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; https://doi.org/10.3389/fpls.2022.1040688.s008
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_6_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s007
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_5_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s006
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_4_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s005
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_3_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; https://doi.org/10.3389/fpls.2022.1040688.s004
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_2_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s003
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_1_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s002
Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; DataSheet_1_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s001
https://doi.org/10.3389/fpls.2022.1040688
Sí
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dc.publisher.none.fl_str_mv Frontiers Media
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spelling Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamianaSaiz-Bonilla, MaríaMartín Merchán, AndreaPallás Benet, VicenteNavarro, José A.Nicotiana benthamianaChloroplastsMitochondriaTranslocon receptorTocTomProtein transportImproved 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.This work was funded by grant PID2020-115571RB-I00 from the Spanish Agencia Estatal de Investigación (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, Innovación y Universidades of Spain, repectively.Frontiers MediaAgencia Estatal de Investigación (España)European CommissionMinisterio de Ciencia, Innovación y Universidades (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2023202320222023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/295536reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-115571RB-I00Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_4_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s012Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_3_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s011Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_2_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s010Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Table_1_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s009Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_7_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; https://doi.org/10.3389/fpls.2022.1040688.s008Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_6_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s007Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_5_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s006Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_4_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s005Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_3_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; https://doi.org/10.3389/fpls.2022.1040688.s004Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_2_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s003Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; Image_1_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.tif [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s002Saiz-Bonilla, María; Martín Merchán, Andrea; Pallás, Vicente; Navarro, José A.; 2022; DataSheet_1_Molecular characterization, targeting and expression analysis of chloroplast and mitochondrion protein import components in Nicotiana benthamiana.docx [Dataset]; Figshare; https://doi.org/10.3389/fpls.2022.1040688.s001https://doi.org/10.3389/fpls.2022.1040688Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2955362026-05-22T06:33:51Z
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