The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis

Comparative Study.

Detalhes bibliográficos
Autores: Fenech, Mario, Amorim-Silva, Vítor, Esteban Del Valle, Alicia, Arnaud, Dominique, Ruiz-López, Noemí, Castillo, Araceli G., Smirnoff, Nicholas, Botella, Miguel A
Tipo de documento: artigo
Estado:Versión aceptada para publicación
Data de publicação:2021
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/374844
Acesso em linha:http://hdl.handle.net/10261/374844
https://api.elsevier.com/content/abstract/scopus_id/85105285938
Access Level:Acceso aberto
Palavra-chave:phosphorylase
ascorbate biosynthesis
GDP-L-galactose
control
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dc.title.none.fl_str_mv The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
title The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
spellingShingle The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
Fenech, Mario
phosphorylase
ascorbate biosynthesis
GDP-L-galactose
control
title_short The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
title_full The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
title_fullStr The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
title_full_unstemmed The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
title_sort The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesis
dc.creator.none.fl_str_mv Fenech, Mario
Amorim-Silva, Vítor
Esteban Del Valle, Alicia
Arnaud, Dominique
Ruiz-López, Noemí
Castillo, Araceli G.
Smirnoff, Nicholas
Botella, Miguel A
author Fenech, Mario
author_facet Fenech, Mario
Amorim-Silva, Vítor
Esteban Del Valle, Alicia
Arnaud, Dominique
Ruiz-López, Noemí
Castillo, Araceli G.
Smirnoff, Nicholas
Botella, Miguel A
author_role author
author2 Amorim-Silva, Vítor
Esteban Del Valle, Alicia
Arnaud, Dominique
Ruiz-López, Noemí
Castillo, Araceli G.
Smirnoff, Nicholas
Botella, Miguel A
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Educación, Cultura y Deporte (España)
Ministerio de Economía, Industria y Competitividad (España)
Ministerio de Ciencia, Innovación y Universidades (España)
Biotechnology and Biological Sciences Research Council (UK)
Universidad de Málaga
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv phosphorylase
ascorbate biosynthesis
GDP-L-galactose
control
topic phosphorylase
ascorbate biosynthesis
GDP-L-galactose
control
description Comparative Study.
publishDate 2021
dc.date.none.fl_str_mv 2021
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
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dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/374844
https://api.elsevier.com/content/abstract/scopus_id/85105285938
url http://hdl.handle.net/10261/374844
https://api.elsevier.com/content/abstract/scopus_id/85105285938
dc.language.none.fl_str_mv Inglés
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info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-82609-R
info:eu-repo/grantAgreement/AEI//AGL2016-75819-C2-1-R
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PGC2018-098789-B-I00
Plant physiology
https://doi.org/10.1093/plphys/kiab010
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Oxford University Press
American Society of Plant Biologists
publisher.none.fl_str_mv Oxford University Press
American Society of Plant Biologists
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
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spelling The role of GDP-l-galactose phosphorylase in the control of ascorbate biosynthesisFenech, MarioAmorim-Silva, VítorEsteban Del Valle, AliciaArnaud, DominiqueRuiz-López, NoemíCastillo, Araceli G.Smirnoff, NicholasBotella, Miguel Aphosphorylaseascorbate biosynthesisGDP-L-galactosecontrolComparative Study.The enzymes involved in l-ascorbate biosynthesis in photosynthetic organisms (the Smirnoff-Wheeler [SW] pathway) are well established. Here, we analyzed their subcellular localizations and potential physical interactions and assessed their role in the control of ascorbate synthesis. Transient expression of C terminal-tagged fusions of SW genes in Nicotiana benthamiana and Arabidopsis thaliana mutants complemented with genomic constructs showed that while GDP-d-mannose epimerase is cytosolic, all the enzymes from GDP-d-mannose pyrophosphorylase (GMP) to l-galactose dehydrogenase (l-GalDH) show a dual cytosolic/nuclear localization. All transgenic lines expressing functional SW protein green fluorescent protein fusions driven by their endogenous promoters showed a high accumulation of the fusion proteins, with the exception of those lines expressing GDP-l-galactose phosphorylase (GGP) protein, which had very low abundance. Transient expression of individual or combinations of SW pathway enzymes in N. benthamiana only increased ascorbate concentration if GGP was included. Although we did not detect direct interaction between the different enzymes of the pathway using yeast-two hybrid analysis, consecutive SW enzymes, as well as the first and last enzymes (GMP and l-GalDH) associated in coimmunoprecipitation studies. This association was supported by gel filtration chromatography, showing the presence of SW proteins in high-molecular weight fractions. Finally, metabolic control analysis incorporating known kinetic characteristics showed that previously reported feedback repression at the GGP step, combined with its relatively low abundance, confers a high-flux control coefficient and rationalizes why manipulation of other enzymes has little effect on ascorbate concentration.This work was supported by a grant from the Spanish Ministerio de Educación, Cultura y Deporte para la formación del Profesorado Universitario (FPU014/01974), I Plan Propio de Investigación, Transferencia y Divulgación Científica de la Universidad de Málaga, The Ministerio de Economía, Industria y Competitividad (RYC-2013-12699), also co-financed by the European Regional Development Fund (BIO2016-81957-REDT, BIO2017-82609-R), the Spanish “Ministerio de Economía, Industria y Competitividad/FEDER” (AGL2016-75819-C2-1-R) and the Spanish Ministerio de Ciencia, Innovación y Universidades (PGC2018-098789-B-I00). N.S. was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) grants BB/G021678/1 and BB/N001311/1Peer reviewedOxford University PressAmerican Society of Plant BiologistsMinisterio de Educación, Cultura y Deporte (España)Ministerio de Economía, Industria y Competitividad (España)Ministerio de Ciencia, Innovación y Universidades (España)Biotechnology and Biological Sciences Research Council (UK)Universidad de MálagaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/374844https://api.elsevier.com/content/abstract/scopus_id/85105285938reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO//RYC-2013-12699info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-82609-Rinfo:eu-repo/grantAgreement/AEI//AGL2016-75819-C2-1-Rinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PGC2018-098789-B-I00Plant physiologyhttps://doi.org/10.1093/plphys/kiab010Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3748442026-05-22T06:33:51Z
score 15.812429

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