Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2014 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFRN |
Texto Completo: | https://repositorio.ufrn.br/jspui/handle/123456789/12573 |
Resumo: | studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins |
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Melo, Julliane Tamara Araújo dehttp://lattes.cnpq.br/3504274193684794http://lattes.cnpq.br/1083882171718362Menck, Carlos Frederico Martinshttp://lattes.cnpq.br/8043136069525312Rocha, Hugo Alexandre de Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799567J8&dataRevisao=nullSouza-pinto, Nadja Cristhina dehttp://lattes.cnpq.br/7088639503480810Lima, Lucymara Fassarela Agnez2014-12-17T14:03:36Z2014-09-052014-12-17T14:03:36Z2014-02-26MELO, Julliane Tamara Araújo de. Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos. 2014. 319 f. Tese (Doutorado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2014.https://repositorio.ufrn.br/jspui/handle/123456789/12573studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteinsA maior parte do nosso conhecimento sobre a via de Reparo de Excisão Nucleotídeos (NER) vem de estudos usando a luz ultravioleta (UV) como fonte de danos no DNA. Contudo, embora os danos no DNA causados pela luz UV sejam relacionados à ocorrência de mutagênese, morte celular e tumorigênese, eles não justificam fenótipos como neurodegeneração e tumorigênese observados em pacientes com síndromes como Xeroderma Pigmentosum (XP) e Síndrome de Cockayne (CS), as quais são associadas à deficiência na via NER. Adicionalmente, evidências mais recentes indicam o envolvimento da via NER no reparo de 8-oxodG, um substrato típico da via de Reparo por Excisão de Bases (BER). Uma vez que a deficiência na via BER resulta em instabilidade genômica, doenças neurodegenerativas e câncer, foi investigado neste trabalho o impacto da deficiência em XPC nas funções da via BER em células humanas. Foram realizadas análises da expressão e da localização celular de APE1, OGG1 e PARP-1, principais enzimas da via BER, em fibroblastos humanos deficientes na via NER. Os resultados demonstraram que os níveis endógenos de APE1, PARP-1 e OGG1 são reduzidos nos fibroblastos deficientes em XPC, os quais foram mais resistentes a diferentes tipos de agentes oxidantes e apresentaram níveis elevados de 8-oxodG quando comparados aos demais fibroblastos deficientes na via NER. Adicionalmente, alterações sutis na localização nuclear e mitocondrial de APE1 foram observadas nos fibroblastos deficientes em XPC. Para confirmar o impacto da deficiência de XPC na regulação da expressão e atividade de APE1 e OGG1, foi construída uma linhagem complementada com XPC. Embora a complementação tenha sido parcial, foi possível observar que os fibroblastos parcialmente complementados com XPC apresentaram níveis maiores de expressão de OGG1 quando comparados aos fibroblastos deficientes em XPC. Os extratos dos fibroblastos parcialmente complementados com XPC também apresentaram uma elevada atividade enzimática de OGG1. Contudo, não foram observadas mudanças na expressão e atividade de APE1 nos fibroblastos parcialmente complementados com XPC. Adicionalmente, foi possível verificar a presença da forma completa de APE1 (37 kDa) e de OGG1-α na mitocôndria dos fibroblastos deficientes em XPC e parcialmente complementados com XPC. Por outro lado, observou-se que a expressão de APE1 e PARP-1 não é alterada no cérebro e fígado de camundongos knockouts para XPC. Contudo, a deficiência em XPC resultou em mudanças na localização celular de APE1 no hipocampo e hipotálamo. Ainda, foi observada a ocorrência de uma interação física entre as proteínas XPC e APE1 em células humanas. Em conclusão, os dados sugerem que a proteína XPC possui um papel na regulação da expressão e da atividade de OGG1 em células humanas e está envolvida na regulação da localização celular de APE1 principalmente em camundongos. Adicionalmente, as respostas celulares dos fibroblastos deficientes na via NER ao estresse oxidativo podem não estar associadas à deficiência na via NER per se, mas podem incluir novas funções das enzimas da via NER na regulação da expressão e localização celular das proteínas da via BERConselho Nacional de Desenvolvimento Científico e Tecnológico2020-01-01application/pdfporUniversidade Federal do Rio Grande do NortePrograma de Pós-Graduação em BioquímicaUFRNBRBioquímica; Biologia MolecularNeurodegeneração. XPC. APE1. OGG1. PARP-1. RegulaçãoNeurodegeneration. XPC. APE1. OGG1. PARP-1. 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dc.title.por.fl_str_mv |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
title |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
spellingShingle |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos Melo, Julliane Tamara Araújo de Neurodegeneração. XPC. APE1. OGG1. PARP-1. Regulação Neurodegeneration. XPC. APE1. OGG1. PARP-1. Regulation CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
title_short |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
title_full |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
title_fullStr |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
title_full_unstemmed |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
title_sort |
Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos |
author |
Melo, Julliane Tamara Araújo de |
author_facet |
Melo, Julliane Tamara Araújo de |
author_role |
author |
dc.contributor.authorID.por.fl_str_mv |
|
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/3504274193684794 |
dc.contributor.advisorID.por.fl_str_mv |
|
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/1083882171718362 |
dc.contributor.referees1.pt_BR.fl_str_mv |
Menck, Carlos Frederico Martins |
dc.contributor.referees1ID.por.fl_str_mv |
|
dc.contributor.referees1Lattes.por.fl_str_mv |
http://lattes.cnpq.br/8043136069525312 |
dc.contributor.referees2.pt_BR.fl_str_mv |
Rocha, Hugo Alexandre de Oliveira |
dc.contributor.referees2ID.por.fl_str_mv |
|
dc.contributor.referees2Lattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799567J8&dataRevisao=null |
dc.contributor.referees3.pt_BR.fl_str_mv |
Souza-pinto, Nadja Cristhina de |
dc.contributor.referees3ID.por.fl_str_mv |
|
dc.contributor.referees3Lattes.por.fl_str_mv |
http://lattes.cnpq.br/7088639503480810 |
dc.contributor.author.fl_str_mv |
Melo, Julliane Tamara Araújo de |
dc.contributor.advisor1.fl_str_mv |
Lima, Lucymara Fassarela Agnez |
contributor_str_mv |
Lima, Lucymara Fassarela Agnez |
dc.subject.por.fl_str_mv |
Neurodegeneração. XPC. APE1. OGG1. PARP-1. Regulação |
topic |
Neurodegeneração. XPC. APE1. OGG1. PARP-1. Regulação Neurodegeneration. XPC. APE1. OGG1. PARP-1. Regulation CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
dc.subject.eng.fl_str_mv |
Neurodegeneration. XPC. APE1. OGG1. PARP-1. Regulation |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA |
description |
studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins |
publishDate |
2014 |
dc.date.accessioned.fl_str_mv |
2014-12-17T14:03:36Z |
dc.date.available.fl_str_mv |
2014-09-05 2014-12-17T14:03:36Z |
dc.date.issued.fl_str_mv |
2014-02-26 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
MELO, Julliane Tamara Araújo de. Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos. 2014. 319 f. Tese (Doutorado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2014. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufrn.br/jspui/handle/123456789/12573 |
identifier_str_mv |
MELO, Julliane Tamara Araújo de. Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos. 2014. 319 f. Tese (Doutorado em Bioquímica; Biologia Molecular) - Universidade Federal do Rio Grande do Norte, Natal, 2014. |
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https://repositorio.ufrn.br/jspui/handle/123456789/12573 |
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por |
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Universidade Federal do Rio Grande do Norte |
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Programa de Pós-Graduação em Bioquímica |
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UFRN |
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BR |
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Bioquímica; Biologia Molecular |
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Universidade Federal do Rio Grande do Norte |
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