Caracterização funcional da proteína Coq10p de Saccharomyces cerevisiae na atividade respiratória da coenzima Q.

Busso, Cleverson

doi:10.11606/T.42.2010.tde-06102010-162442

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Doctoral Thesis

DOI

https://doi.org/10.11606/T.42.2010.tde-06102010-162442

Document

Doctoral Thesis

Author

Busso, Cleverson (Catálogo USP)

Full name

Cleverson Busso

Institute/School/College

Instituto de Ciências Biomédicas

Knowledge Area

Microbiology

Date of Defense

2010-08-25

Published

São Paulo, 2010

Supervisor

Barros, Mário Henrique de (Catálogo USP)
Ferreira Júnior, José Ribamar dos Santos - (Co-supervisor) (Catálogo USP)

Committee

Barros, Mário Henrique de (President)
Maia, Ivan de Godoy
Nobrega, Francisco Gorgonio da
Oliveira, Carla Columbano de
Soares Netto, Luis Eduardo

Title in Portuguese

Caracterização funcional da proteína Coq10p de Saccharomyces cerevisiae na atividade respiratória da coenzima Q.

Keywords in Portuguese

Biologia molecular
Estresse oxidativo
Leveduras
Microbiologia
Mitocôndrias
Mutagênese

Abstract in Portuguese

A coenzima Q transporta elétrons das NADH e succinato desidrogenase para o complexo bc1 da cadeia respiratória mitocondrial. Há dez produtos gênicos conhecidos em seu metabolismo; este trabalho foca o último a ser descrito: COQ10. Considerando a presença de coenzima Q no interior de células contendo este gene inativo, verificamos a atividade respiratória da coenzima Q em mutantes coq10. Através de inibidores do ciclo da coenzima Q no interior do complexo III da cadeia respiratória mitocondrial (antimicina A e mixotiazol) na linhagem coq10, notou-se sua resposta a antimicina A, mas não a mixotiazol, sugerindo a existência de semiquinonas, que também foram confirmadas em testes de ressonância paramagnética eletrônica. O possível transporte e distribuição da coenzima Q por Coq10p foi estudado através de mutações sítio-dirigidas em posições críticas de um túnel de ligação existente em sua estrutura modelar, bem como por análise de super-expressão da proteína. Este trabalho confirma o papel central de Coq10p no metabolismo respiratório.

Title in English

Functional characterization of the Coq10p protein of Saccharomyces cerevisiae in coenzyme Q respiratory activity.

Keywords in English

Microbiology
Mitochondria
Molecular biology
Mutagenesis
Oxidative stress
Yeast

Abstract in English

Coenzyme Q transports electrons from NADH and succinate dehydrogenase to the bc1 complex of the mitochondrial respiratory chain. Ten gene products are involved in coenzyme Q metabolism. This work focuses on the last gene to be described: COQ10. Considering the presence of coenzyme Q in the interior of cells containing this gene inactive, we analyzed the respiratory activity of coenzyme Q in coq10 mutants. By using Q cycle inhibitors, in the interior of the complex III of mitochondrial respiratory chain (antimycin A and mixotiazol), in coq10 lineage, it was noted the response to antimycin A, but not mixotiazol, suggesting the existence of semiquinones, which were also confirmed in tests of electron paramagnetic resonance. The possible transmission and distribution of coenzyme Q by Coq10p was studied by site-directed mutagenesis at critical positions of a tunnel present in the molecular model structure, as well as analysis of over-expression of the protein. This study confirms the central role of Coq10p in respiratory metabolism.

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Publishing Date

2010-10-28

Derived works

WARNING: The material described below relates to works resulting from this thesis or dissertation. The contents of these works are the author's responsibility.

Busso, Cleverson, et al. Saccharomyces cerevisiae coq10 null mutants are responsive to antimycin A [doi:10.1111/j.1742-4658.2010.07862.x]. The FEBS Journal [online], 2010, vol. 277, p. 4530-4538.
Busso, Cleverson, et al. Site-directed mutagenesis and structural modeling of Coq10p indicate the presence of a tunnel for coenzyme Q6 binding [doi:10.1016/j.febslet.2010.03.024]. FEBS Letters [online], 2010, vol. 584, p. 1609-1614.
Gomes, Fernando, et al. nde1 deletion improves mitochondrial DNA maintenance in Saccharomyces cerevisiae coenzyme Q mutants [doi:10.1042/BJ20121432]. Biochemical Journal (London. 1984) [online], 2012, vol. 449, p. 595-603.
LAGIERTOURENNE, C, et al. ADCK3, an Ancestral Kinase, Is Mutated in a Form of Recessive Ataxia Associated with Coenzyme Q10 Deficiency [doi:10.1016/j.ajhg.2007.12.024]. American Journal of Human Genetics [online], 2008, vol. 82, p. 661-672.
Zampol, Mariana A., et al. Over-expression of COQ10 in Saccharomyces cerevisiae inhibits mitochondrial respiration [doi:10.1016/j.bbrc.2010.09.118]. Biochemical and Biophysical Research Communications [online], 2010, vol. 402, p. 82-87.
BUSSO, C., et al. Estudo do estresse oxidativo em mutantes de Saccharomyces cerevisiae deficientes em coenzima Q6. In 53º Congresso Brasileiro de Genética,, Águas de Lindóia/SP, 2007. Caderno de Resumos.Ribeirão Preto : SBBG, 2007. Resumo.
BUSSO, C, Paulela, J. A., e BARROS, M. H. Estudo Funcional da Proteína COQ7 em Mutantes Sensíveis a Temperatura de Saccharomyces cerevisiae. In 26 Congresso Brasileiro de Microbiologia, Foz do Iguaçu, 2011. Microbiologia em Foco.Sao Paulo : Vox Editora, 2011. Resumo.
BUSSO, C., et al. New evidences of Coq10p role in coenzyme Q distribuition. In 21st IUBMB and 12th FAOBMB International Congress of Biochemistry and Molecular Biology, 2009, Shangai, China, 2009. Abstracts - Biomolecules for Quality of Life, 2009.., 2009. Abstract.
BUSSO, C., et al. Saccharomyces cerevisiae coq10 mutants have a compromised q-cycle. In XXXIX Annual Meeting of SBBq, Foz do Iguaçu/PR, 2010. SBBq-Program of the XXXIX Annual Meeting.São Paulo : SBBq, 2010. Abstract.
BUSSO, C., et al. Sequence analysis and site direct mutagenesis of S. cerevisiae COQ10 are consistent with the existence of START domain. In 21st IUBMB and 12th FAOBMB International Congress of Biochemistry and Molecular Biology, 2009, Shangai, China, 2009. Biomolecules for Quality of Life, 2009., 2009. Abstract.
BUSSO, C., et al. Site Directed Mutagenesis and Structure Model Corroborate the Presence of a Coenzyme Q Binding Domain in Saccharomyces cerevisiae COQ10. In XXXVII Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology (SBBq), Águas de Lindóia, 2008. SBBq-Program of the XXXVII Annual Meeting - 2008.São Paulo : SBBq, 2008. Abstract.
Paulela, J. A., et al. Caracterização de mutantes coq7 sensíveis a temperatura e análise estrutural das sequencias alteradas da proteína. In XXI ALAM Congresso Latinoamericano de Microbiologia, Santos, 2012. XXI ALAM Congresso Latinoamericano de Microbiologia.São Paulo : SBM, 2012. Resumo.
ZAMPOL, M. A., et al. Regulated Expression of COQ10 in S. Cerevisiae Coenzyme Q6 Mutants. In XXXVII Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology (SBBq), Águas de Lindóia/SP, 2008. SBBq Program of the XXXVII Annual Meeting 2008.São Paulo : SBBq, 2008. Abstract.