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Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis()
Coq5 catalyzes the only C-methylation involved in the biosynthesis of coenzyme Q (Q or ubiquinone) in humans and yeast Saccharomyces cerevisiae. As one of eleven polypeptides required for Q production in yeast, Coq5 has also been shown to assemble with the multi-subunit complex termed the CoQ-syntho...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4331671/ https://www.ncbi.nlm.nih.gov/pubmed/25152161 http://dx.doi.org/10.1016/j.bbalip.2014.08.007 |
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author | Nguyen, Theresa P.T. Casarin, Alberto Desbats, Maria Andrea Doimo, Mara Trevisson, Eva Santos-Ocaña, Carlos Navas, Placido Clarke, Catherine F. Salviati, Leonardo |
author_facet | Nguyen, Theresa P.T. Casarin, Alberto Desbats, Maria Andrea Doimo, Mara Trevisson, Eva Santos-Ocaña, Carlos Navas, Placido Clarke, Catherine F. Salviati, Leonardo |
author_sort | Nguyen, Theresa P.T. |
collection | PubMed |
description | Coq5 catalyzes the only C-methylation involved in the biosynthesis of coenzyme Q (Q or ubiquinone) in humans and yeast Saccharomyces cerevisiae. As one of eleven polypeptides required for Q production in yeast, Coq5 has also been shown to assemble with the multi-subunit complex termed the CoQ-synthome. In humans, mutations in several COQ genes cause primary Q deficiency, and a decrease in Q biosynthesis is associated with mitochondrial, cardiovascular, kidney and neurodegenerative diseases. In this study, we characterize the human COQ5 polypeptide and examine its complementation of yeast coq5 point and null mutants. We show that human COQ5 RNA is expressed in all tissues and that the COQ5 polypeptide is associated with the mitochondrial inner membrane on the matrix side. Previous work in yeast has shown that point mutations within or adjacent to conserved COQ5 methyltransferase motifs result in a loss of Coq5 function but not Coq5 steady state levels. Here, we show that stabilization of the CoQ-synthome within coq5 point mutants or by over-expression of COQ8 in coq5 null mutants permits the human COQ5 homolog to partially restore coq5 mutant growth on respiratory media and Q(6) content. Immunoblotting against the human COQ5 polypeptide in isolated yeast mitochondria shows that the human Coq5 polypeptide migrates in two-dimensional blue-native/SDS-PAGE at the same high molecular mass as other yeast Coq proteins. The results presented suggest that human and Escherichia coli Coq5 homologs expressed in yeast retain C-methyltransferase activity but are capable of rescuing the coq5 yeast mutants only when the CoQ-synthome is assembled. |
format | Online Article Text |
id | pubmed-4331671 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-43316712015-03-03 Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() Nguyen, Theresa P.T. Casarin, Alberto Desbats, Maria Andrea Doimo, Mara Trevisson, Eva Santos-Ocaña, Carlos Navas, Placido Clarke, Catherine F. Salviati, Leonardo Biochim Biophys Acta Mol Cell Biol Lipids Article Coq5 catalyzes the only C-methylation involved in the biosynthesis of coenzyme Q (Q or ubiquinone) in humans and yeast Saccharomyces cerevisiae. As one of eleven polypeptides required for Q production in yeast, Coq5 has also been shown to assemble with the multi-subunit complex termed the CoQ-synthome. In humans, mutations in several COQ genes cause primary Q deficiency, and a decrease in Q biosynthesis is associated with mitochondrial, cardiovascular, kidney and neurodegenerative diseases. In this study, we characterize the human COQ5 polypeptide and examine its complementation of yeast coq5 point and null mutants. We show that human COQ5 RNA is expressed in all tissues and that the COQ5 polypeptide is associated with the mitochondrial inner membrane on the matrix side. Previous work in yeast has shown that point mutations within or adjacent to conserved COQ5 methyltransferase motifs result in a loss of Coq5 function but not Coq5 steady state levels. Here, we show that stabilization of the CoQ-synthome within coq5 point mutants or by over-expression of COQ8 in coq5 null mutants permits the human COQ5 homolog to partially restore coq5 mutant growth on respiratory media and Q(6) content. Immunoblotting against the human COQ5 polypeptide in isolated yeast mitochondria shows that the human Coq5 polypeptide migrates in two-dimensional blue-native/SDS-PAGE at the same high molecular mass as other yeast Coq proteins. The results presented suggest that human and Escherichia coli Coq5 homologs expressed in yeast retain C-methyltransferase activity but are capable of rescuing the coq5 yeast mutants only when the CoQ-synthome is assembled. Elsevier 2014-11 /pmc/articles/PMC4331671/ /pubmed/25152161 http://dx.doi.org/10.1016/j.bbalip.2014.08.007 Text en © 2014 The Authors. Published by Elsevier B.V. https://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). |
spellingShingle | Article Nguyen, Theresa P.T. Casarin, Alberto Desbats, Maria Andrea Doimo, Mara Trevisson, Eva Santos-Ocaña, Carlos Navas, Placido Clarke, Catherine F. Salviati, Leonardo Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() |
title | Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() |
title_full | Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() |
title_fullStr | Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() |
title_full_unstemmed | Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() |
title_short | Molecular characterization of the human COQ5 C-methyltransferase in coenzyme Q(10) biosynthesis() |
title_sort | molecular characterization of the human coq5 c-methyltransferase in coenzyme q(10) biosynthesis() |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4331671/ https://www.ncbi.nlm.nih.gov/pubmed/25152161 http://dx.doi.org/10.1016/j.bbalip.2014.08.007 |
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