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Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.

Detalhes bibliográficos
Ano de defesa: 2023
Autor(a) principal: Souza, Paulo Vinicius Leite de
Orientador(a): Daloso, Danilo de Menezes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/71966
Resumo: Thioredoxins (TRXs) consist of a family of oxi-redox proteins capable of (de)activating enzymes, being an important post-translational mechanism for metabolic regulation. Plant TRXs are found in different cell compartments, such as chloroplast, mitochondria, and cytosol. TRXs are activated by specific TRX reductases (TRs), according to their subcellular location. For instance, chloroplasts contain two TRs, named FTR (ferredoxin-dependent TR) and NTRC (NADPH- dependent TR C), which receives reducing power from ferredoxin and NADPH, respectivelly. On the other hand, plant cells contain two other NTRs (NTRA and NTRB) that are mainly located in cytosol and mitochondria. NTRA/B are thus the main TRs responsible for reducing the non- chloroplastic TRXs. Previous works showed that the single ntrc mutant and the double ntrab mutant have reduced growth, but are still viable, i.e. they can complete the full life cycle and produce viable seeds. However, plants lacking all NTRs remained to be investigated, raising the question on whether the plant NTR system is essential for plant growth and development. In paralell, previous works suggested that the mitocondrial NTR/TRX system can regulate the interplay between carbon and nitrogen metabolisms, but how redox-mediated mechanisms regulate these metabolisms remained to be deeply investigated. Aiming to address these questions, this thesis was divided into three parts. We first reviewed how TRX-mediated mechanisms regulate the primary metabolism, especially the (photo)respiratory metabolism. The second and third chapters involve the characterization of Arabidopsis thaliana L mutants lacking different NTRs and TRXs. The second chapter describes the unprecedented characterization of the triple ntrabc mutant, which lacks all NTRs (NTR A, B and C), whilst the third involves experiments using mutants lacking TRX h2, TRX o1 or NTRA/B aiming to investigate how the NTR/TRX system regulates glutamine synthetase (GS) and the fluxes throughout the GS/GOGAT (glutamate synthase) cycle. Our results showed that the triple ntrabc mutant showed a leaf pale green phenotype with strong reduction in growth and substantial metabolic changes. Despite this, the ntrabc remained viable and was able to complete the full developmental cycle, including the production of viable seeds. These results suggest that the NTR system is highly important for plant growth, but not essential for plant development. Furthermore, our results indicate that the mitochondrial NTR/TRX system is key for the regulation of the redox status of GS and the metabolic fluxes thorughout the GS/GOGAT cycle, which is important for plant high light stress acclimation. Thus, our works provide important and unprecendented informations regarding the regulation of primary metabolism mediated by NTRs and TRXs.
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spelling Souza, Paulo Vinicius Leite deDaloso, Danilo de Menezes2023-04-28T15:03:48Z2023-04-28T15:03:48Z2023SOUZA, Paulo Vinicius Leite de. Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.. 2023. 153 f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2023.http://www.repositorio.ufc.br/handle/riufc/71966Thioredoxins (TRXs) consist of a family of oxi-redox proteins capable of (de)activating enzymes, being an important post-translational mechanism for metabolic regulation. Plant TRXs are found in different cell compartments, such as chloroplast, mitochondria, and cytosol. TRXs are activated by specific TRX reductases (TRs), according to their subcellular location. For instance, chloroplasts contain two TRs, named FTR (ferredoxin-dependent TR) and NTRC (NADPH- dependent TR C), which receives reducing power from ferredoxin and NADPH, respectivelly. On the other hand, plant cells contain two other NTRs (NTRA and NTRB) that are mainly located in cytosol and mitochondria. NTRA/B are thus the main TRs responsible for reducing the non- chloroplastic TRXs. Previous works showed that the single ntrc mutant and the double ntrab mutant have reduced growth, but are still viable, i.e. they can complete the full life cycle and produce viable seeds. However, plants lacking all NTRs remained to be investigated, raising the question on whether the plant NTR system is essential for plant growth and development. In paralell, previous works suggested that the mitocondrial NTR/TRX system can regulate the interplay between carbon and nitrogen metabolisms, but how redox-mediated mechanisms regulate these metabolisms remained to be deeply investigated. Aiming to address these questions, this thesis was divided into three parts. We first reviewed how TRX-mediated mechanisms regulate the primary metabolism, especially the (photo)respiratory metabolism. The second and third chapters involve the characterization of Arabidopsis thaliana L mutants lacking different NTRs and TRXs. The second chapter describes the unprecedented characterization of the triple ntrabc mutant, which lacks all NTRs (NTR A, B and C), whilst the third involves experiments using mutants lacking TRX h2, TRX o1 or NTRA/B aiming to investigate how the NTR/TRX system regulates glutamine synthetase (GS) and the fluxes throughout the GS/GOGAT (glutamate synthase) cycle. Our results showed that the triple ntrabc mutant showed a leaf pale green phenotype with strong reduction in growth and substantial metabolic changes. Despite this, the ntrabc remained viable and was able to complete the full developmental cycle, including the production of viable seeds. These results suggest that the NTR system is highly important for plant growth, but not essential for plant development. Furthermore, our results indicate that the mitochondrial NTR/TRX system is key for the regulation of the redox status of GS and the metabolic fluxes thorughout the GS/GOGAT cycle, which is important for plant high light stress acclimation. Thus, our works provide important and unprecendented informations regarding the regulation of primary metabolism mediated by NTRs and TRXs.As tioredoxinas (TRXs) constituem uma família de proteínas oxi-redox capazes de (des)ativar enzimas, sendo um importante mecanismo pós-traducional para a regulação do fluxo metabólico. As TRXs vegetais estão espalhados em diferentes compartimentos celulares, como cloroplastos, mitocôndrias e citosol. As TRXs são ativadas por TRX redutases (TRs). Os cloroplastos possuem dois TRs, a saber, FTR (TR dependente de ferredoxina) e NTRC (TR dependente de NADPH), que recebe poder redutor da ferredoxina e/ou NADPH. Por outro lado, as células vegetais contêm duas outras NTRs fora do cloroplasto, ou seja, as isoformas NTRA e NTRB que estão localizadas principalmente no citosol e nas mitocôndrias. As NTRA/B são, portanto, as principais TRs responsáveis pela redução dos TRXs não plastidiais. Trabalhos anteriores mostraram que o mutante nocaute para ntrc e ntrab apresentam forte decréscimo no crescimento, mas ainda são viáveis, ou seja, podem completar o ciclo de vida completo e produzir sementes viáveis. No entanto, as plantas que carecem de todos as NTRs ainda precisam serem investigadas, abrindo a questão sobre se o sistema NTR de plantas é essencial para o crescimento e desenvolvimento destes organismos. Paralelamente, trabalhos anteriores sugeriram que o sistema mitocondrial NTR/TRX pode regular a interação entre os metabolismos de carbono e nitrogênio, mas como os mecanismos mediados por redox regulam esses metabolismos ainda precisam ser profundamente investigados. Com o objetivo de responder a essas questões, esta tese foi dividida em três partes. Primeiro revisamos como os mecanismos mediados por TRX regulam o metabolismo primário, especialmente o metabolismo (foto)respiratório. O segundo e o terceiro capítulos envolvem a caracterização de mutantes da planta modelo Arabidopsis thaliana L. O segundo capítulo descreve a caracterização inédita do mutante triplo ntrabc, que carece de todos os NTRs (NTR A, B e C), enquanto o terceiro envolve experimentos usando mutantes sem TRX h2, TRX o1 ou NTRA/B com o objetivo de investigar como o sistema NTR/TRX regula a glutamina sintetase (GS) e os fluxos ao longo do ciclo GS/GOGAT (glutamato sintase). Nossos resultados mostraram que o mutante triplo ntrabc apresentou um fenótipo de folha verde pálida com forte redução no crescimento e alterações metabólicas substanciais. Apesar disso, o ntrabc manteve-se viável e conseguiu completar todo o ciclo de desenvolvimento, incluindo a produção de sementes viáveis. Nossos resultados sugerem que o sistema NTR é altamente importante para o crescimento da planta, mas não essencial para o desenvolvimento da planta. Além disso, nossos resultados indicam ainda que o sistema NTR/TRX mitocondrial é a chave para a regulação do status redox de GS e os fluxos metabólicos ao longo do ciclo GS/GOGAT, o que é importante para a aclimatação da planta ao estresse de alta luz. Assim, nossos trabalhos fornecem informações importantes e inéditas sobre a regulação do metabolismo primário mediado por NTRs e TRXs.TiorredoxinasNADPH - Tioredoxina redutaseMetabolismo primárioMetabolismo do nitrogênioEstresse de alta luzMetabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2023_tese_pvlsouza.pdf2023_tese_pvlsouza.pdfapplication/pdf8604753http://repositorio.ufc.br/bitstream/riufc/71966/3/2023_tese_pvlsouza.pdfdc81ce6519d984f01bd65ff7881ef740MD53LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/71966/4/license.txt8a4605be74aa9ea9d79846c1fba20a33MD54riufc/719662023-04-28 12:03:49.279oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2023-04-28T15:03:49Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
dc.title.en.pt_BR.fl_str_mv Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
title Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
spellingShingle Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
Souza, Paulo Vinicius Leite de
Tiorredoxinas
NADPH - Tioredoxina redutase
Metabolismo primário
Metabolismo do nitrogênio
Estresse de alta luz
title_short Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
title_full Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
title_fullStr Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
title_full_unstemmed Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
title_sort Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.
author Souza, Paulo Vinicius Leite de
author_facet Souza, Paulo Vinicius Leite de
author_role author
dc.contributor.author.fl_str_mv Souza, Paulo Vinicius Leite de
dc.contributor.advisor1.fl_str_mv Daloso, Danilo de Menezes
contributor_str_mv Daloso, Danilo de Menezes
dc.subject.por.fl_str_mv Tiorredoxinas
NADPH - Tioredoxina redutase
Metabolismo primário
Metabolismo do nitrogênio
Estresse de alta luz
topic Tiorredoxinas
NADPH - Tioredoxina redutase
Metabolismo primário
Metabolismo do nitrogênio
Estresse de alta luz
description Thioredoxins (TRXs) consist of a family of oxi-redox proteins capable of (de)activating enzymes, being an important post-translational mechanism for metabolic regulation. Plant TRXs are found in different cell compartments, such as chloroplast, mitochondria, and cytosol. TRXs are activated by specific TRX reductases (TRs), according to their subcellular location. For instance, chloroplasts contain two TRs, named FTR (ferredoxin-dependent TR) and NTRC (NADPH- dependent TR C), which receives reducing power from ferredoxin and NADPH, respectivelly. On the other hand, plant cells contain two other NTRs (NTRA and NTRB) that are mainly located in cytosol and mitochondria. NTRA/B are thus the main TRs responsible for reducing the non- chloroplastic TRXs. Previous works showed that the single ntrc mutant and the double ntrab mutant have reduced growth, but are still viable, i.e. they can complete the full life cycle and produce viable seeds. However, plants lacking all NTRs remained to be investigated, raising the question on whether the plant NTR system is essential for plant growth and development. In paralell, previous works suggested that the mitocondrial NTR/TRX system can regulate the interplay between carbon and nitrogen metabolisms, but how redox-mediated mechanisms regulate these metabolisms remained to be deeply investigated. Aiming to address these questions, this thesis was divided into three parts. We first reviewed how TRX-mediated mechanisms regulate the primary metabolism, especially the (photo)respiratory metabolism. The second and third chapters involve the characterization of Arabidopsis thaliana L mutants lacking different NTRs and TRXs. The second chapter describes the unprecedented characterization of the triple ntrabc mutant, which lacks all NTRs (NTR A, B and C), whilst the third involves experiments using mutants lacking TRX h2, TRX o1 or NTRA/B aiming to investigate how the NTR/TRX system regulates glutamine synthetase (GS) and the fluxes throughout the GS/GOGAT (glutamate synthase) cycle. Our results showed that the triple ntrabc mutant showed a leaf pale green phenotype with strong reduction in growth and substantial metabolic changes. Despite this, the ntrabc remained viable and was able to complete the full developmental cycle, including the production of viable seeds. These results suggest that the NTR system is highly important for plant growth, but not essential for plant development. Furthermore, our results indicate that the mitochondrial NTR/TRX system is key for the regulation of the redox status of GS and the metabolic fluxes thorughout the GS/GOGAT cycle, which is important for plant high light stress acclimation. Thus, our works provide important and unprecendented informations regarding the regulation of primary metabolism mediated by NTRs and TRXs.
publishDate 2023
dc.date.accessioned.fl_str_mv 2023-04-28T15:03:48Z
dc.date.available.fl_str_mv 2023-04-28T15:03:48Z
dc.date.issued.fl_str_mv 2023
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.identifier.citation.fl_str_mv SOUZA, Paulo Vinicius Leite de. Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.. 2023. 153 f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2023.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/71966
identifier_str_mv SOUZA, Paulo Vinicius Leite de. Metabolic regulation mediated by thioredoxins and NADPH-dependent thioredoxin reductases in Arabidopsis thaliana L.. 2023. 153 f. Tese (Doutorado em Bioquímica) - Universidade Federal do Ceará, Fortaleza, 2023.
url http://www.repositorio.ufc.br/handle/riufc/71966
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