FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2943082710> ?p ?o ?g. }

• W2943082710 endingPage "1310" @default.
• W2943082710 startingPage "1293" @default.
• W2943082710 abstract "Coenzyme Q (CoQ or ubiquinone) serves as an essential redox-active lipid in respiratory electron and proton transport during cellular energy metabolism. CoQ also functions as a membrane-localized antioxidant protecting cells against lipid peroxidation. CoQ deficiency is associated with multiple human diseases; CoQ10 supplementation in particular has noted cardioprotective benefits. In Saccharomyces cerevisiae, Coq10, a putative START domain protein, is believed to chaperone CoQ to sites where it functions. Yeast coq10 deletion mutants (coq10Δ) synthesize CoQ inefficiently during log phase growth and are respiratory defective and sensitive to oxidative stress. Humans have two orthologs of yeast COQ10, COQ10A and COQ10B. Here, we tested the human co-orthologs for their ability to rescue the yeast mutant. We showed that expression of either human ortholog, COQ10A or COQ10B, rescues yeast coq10Δ mutant phenotypes, restoring the function of respiratory-dependent growth on a nonfermentable carbon source and sensitivity to oxidative stress induced by treatment with PUFAs. These effects indicate a strong functional conservation of Coq10 across different organisms. However, neither COQ10A nor COQ10B restored CoQ biosynthesis when expressed in the yeast coq10Δ mutant. The involvement of yeast Coq10 in CoQ biosynthesis may rely on its interactions with another protein, possibly Coq11, which is not found in humans. Coexpression analyses of yeast COQ10 and human COQ10A and COQ10B provide additional insights to functions of these START domain proteins and their potential roles in other biologic pathways. Coenzyme Q (CoQ or ubiquinone) serves as an essential redox-active lipid in respiratory electron and proton transport during cellular energy metabolism. CoQ also functions as a membrane-localized antioxidant protecting cells against lipid peroxidation. CoQ deficiency is associated with multiple human diseases; CoQ10 supplementation in particular has noted cardioprotective benefits. In Saccharomyces cerevisiae, Coq10, a putative START domain protein, is believed to chaperone CoQ to sites where it functions. Yeast coq10 deletion mutants (coq10Δ) synthesize CoQ inefficiently during log phase growth and are respiratory defective and sensitive to oxidative stress. Humans have two orthologs of yeast COQ10, COQ10A and COQ10B. Here, we tested the human co-orthologs for their ability to rescue the yeast mutant. We showed that expression of either human ortholog, COQ10A or COQ10B, rescues yeast coq10Δ mutant phenotypes, restoring the function of respiratory-dependent growth on a nonfermentable carbon source and sensitivity to oxidative stress induced by treatment with PUFAs. These effects indicate a strong functional conservation of Coq10 across different organisms. However, neither COQ10A nor COQ10B restored CoQ biosynthesis when expressed in the yeast coq10Δ mutant. The involvement of yeast Coq10 in CoQ biosynthesis may rely on its interactions with another protein, possibly Coq11, which is not found in humans. Coexpression analyses of yeast COQ10 and human COQ10A and COQ10B provide additional insights to functions of these START domain proteins and their potential roles in other biologic pathways. Coenzyme Q (CoQ) is a lipid composed of a fully substituted redox-active benzoquinone ring attached to a long polyisoprenoid chain. The polyisoprenoid chain of CoQn, with n ≥ 6 isoprene units, anchors CoQ at the mid-plane of the membrane phospholipid bilayers. The reversible reduction and oxidation of CoQ and CoQH2 enables the transport of electrons and protons necessary for cellular respiration. CoQ also serves as an important electron acceptor for enzymes involved in fatty acid β-oxidation, oxidation of proline and sulfide, and pyrimidine biosynthesis (1Alcázar-Fabra M. Trevisson E. Brea-Calvo G. Clinical syndromes associated with coenzyme Q10 deficiency.Essays Biochem. 2018; 62: 377-398Crossref PubMed Scopus (32) Google Scholar, 2Turunen M. Olsson J. Dallner G. Metabolism and function of coenzyme Q.Biochim. Biophys. Acta. 2004; 1660: 171-199Crossref PubMed Scopus (691) Google Scholar, 3Desbats M.A. Lunardi G. Doimo M. Trevisson E. Salviati L. Genetic bases and clinical manifestations of coenzyme Q10 (CoQ10) deficiency.J. Inherit. Metab. Dis. 2015; 38: 145-156Crossref PubMed Scopus (0) Google Scholar). The reduced or hydroquinone form of CoQH2 serves as a chain-terminating antioxidant that slows lipid peroxidation (2Turunen M. Olsson J. Dallner G. Metabolism and function of coenzyme Q.Biochim. Biophys. Acta. 2004; 1660: 171-199Crossref PubMed Scopus (691) Google Scholar). Although CoQ exists in most biological membranes, its synthesis occurs exclusively inside the mitochondria in eukaryotes, or in the cytosol in Escherichia coli, catalyzed by a cohort of enzymes, many of which are organized in a complex known as the CoQ synthome (also known as complex Q) in eukaryotes, or the Ubi metabolon in E. coli (4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar, 5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar, 6Hajj Chehade M. Pelosi L. Fyfe C.D. Loiseau L. Rascalou B. Brugière S. Kazemzadeh K. Vo C-D-T. Ciccone L. Aussel L. et al.A soluble metabolon synthesizes the isoprenoid lipid ubiquinone.Cell Chem. Biol. 2019; 26: 482-492.e7Abstract Full Text Full Text PDF PubMed Scopus (11) Google Scholar). In Saccharomyces cerevisiae, currently known members of the CoQ synthome consist of Coq3–Coq9 and Coq11 (4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar, 5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar). Together, they modify the quinone head group through a series of methylation (Coq3 and Coq5), deamination (Coq6, Coq9), and hydroxylation (Coq6, Coq7, Coq9) reactions (4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar, 5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar). The definitive functions of the remaining members of the CoQ synthome (Coq4, Coq8, and Coq11) are yet to be fully characterized. Attachment of the polyisoprenoid chain to the aromatic ring precursor precedes the ring modification steps. Coq1, a hexaprenyl pyrophosphate synthetase, condenses farnesyl pyrophosphate with three molecules of isopentenyl pyrophosphate to form hexaprenyldiphosphate, which is transferred to the 4-hydroxybenzoic acid (4HB) or para-aminobenzoic acid (pABA) ring at the C3 position by Coq2 (4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar, 5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar). The number of isoprene units (n) in the polyisoprenoid chain of CoQn varies between organisms, as determined by the specific polyprenyl diphosphate synthase (7Okada K. Suzuki K. Kamiya Y. Zhu X. Fujisaki S. Nishimura Y. Nishino T. Nakagawa T. Kawamukai M. Matsuda H. Polyprenyl diphosphate synthase essentially defines the length of the side chain of ubiquinone.Biochim. Biophys. Acta. 1996; 1302: 217-223Crossref PubMed Scopus (98) Google Scholar), and consists of six isoprene units in S. cerevisiae (CoQ6), eight isoprene units in E. coli (CoQ8), and predominantly ten isoprene units in Schizosaccharomyces pombe and humans (CoQ10) (8Kawamukai M. Biosynthesis of coenzyme Q in eukaryotes.Biosci. Biotechnol. Biochem. 2016; 80: 23-33Crossref PubMed Scopus (55) Google Scholar). Each of the yeast coq1Δ–coq9Δ mutants shows complete abolishment of CoQ6 biosynthesis and fails to respire (5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar). Their defects in respiration can be readily restored by exogenous supplementation with CoQ6 (5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar). The coq10Δ mutant is unusual among the yeast coq mutants because it produces wild-type content of CoQ6 at stationary phase, yet its de novo synthesis of CoQ6 during log phase is inefficient (9Allan C.M. Hill S. Morvaridi S. Saiki R. Johnson J.S. Liau W.S. Hirano K. Kawashima T. Ji Z. Loo J.A. et al.A conserved START domain coenzyme Q-binding polypeptide is required for efficient Q biosynthesis, respiratory electron transport, and antioxidant function in Saccharomyces cerevisiae.Biochim. Biophys. Acta. 2013; 1831: 776-791Crossref PubMed Scopus (23) Google Scholar, 10Barros M.H. Johnson A. Gin P. Marbois B.N. Clarke C.F. Tzagoloff A. The Saccharomyces cerevisiae COQ10 gene encodes a START domain protein required for function of coenzyme Q in respiration.J. Biol. Chem. 2005; 280: 42627-42635Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar). Despite having normal or nearly normal steady state levels of CoQ6, the coq10Δ mutant displays a respiratory-deficient phenotype shown by anemic growth on medium containing a nonfermentable carbon source and decreased NADH and succinate oxidase activities (10Barros M.H. Johnson A. Gin P. Marbois B.N. Clarke C.F. Tzagoloff A. The Saccharomyces cerevisiae COQ10 gene encodes a START domain protein required for function of coenzyme Q in respiration.J. Biol. Chem. 2005; 280: 42627-42635Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar). In addition, the coq10Δ mutant is sensitive to lipid peroxidation induced by exogenously added PUFAs (9Allan C.M. Hill S. Morvaridi S. Saiki R. Johnson J.S. Liau W.S. Hirano K. Kawashima T. Ji Z. Loo J.A. et al.A conserved START domain coenzyme Q-binding polypeptide is required for efficient Q biosynthesis, respiratory electron transport, and antioxidant function in Saccharomyces cerevisiae.Biochim. Biophys. Acta. 2013; 1831: 776-791Crossref PubMed Scopus (23) Google Scholar). Thus, the CoQ6 present in the coq10Δ mutant is not utilized efficiently for either respiration or for its function as an antioxidant. The NMR structure of CC1736, a Coq10 ortholog in Caulobacter crescentus, identified it as a member of the steroidogenic acute regulatory protein-related lipid transfer (START) domain superfamily (11Shen Y. Goldsmith-Fischman S. Atreya H.S. Acton T. Ma L. Xiao R. Honig B. Montelione G.T. Szyperski T. NMR structure of the 18 kDa protein CC1736 from Caulobacter crescentus identifies a member of the START domain superfamily and suggests residues mediating substrate specificity.Proteins. 2005; 58: 747-750Crossref PubMed Scopus (17) Google Scholar). This family includes proteins that bind polycyclic compounds, such as cholesterol and polyketides, in a signature hydrophobic cavity (11Shen Y. Goldsmith-Fischman S. Atreya H.S. Acton T. Ma L. Xiao R. Honig B. Montelione G.T. Szyperski T. NMR structure of the 18 kDa protein CC1736 from Caulobacter crescentus identifies a member of the START domain superfamily and suggests residues mediating substrate specificity.Proteins. 2005; 58: 747-750Crossref PubMed Scopus (17) Google Scholar). The START domain typically spans ∼210 residues (12Ponting C.P. Aravind L. START: a lipid-binding domain in StAR, HD-ZIP and signalling proteins.Trends Biochem. Sci. 1999; 24: 130-132Abstract Full Text Full Text PDF PubMed Scopus (319) Google Scholar) and folds into a helix-grip structure consisting of antiparallel β-sheets flanked by one α-helix on each side (13Iyer L.M. Koonin E.V. Aravind L. Adaptations of the helix-grip fold for ligand binding and catalysis in the START domain superfamily.Proteins. 2001; 43: 134-144Crossref PubMed Scopus (174) Google Scholar). START domain-containing proteins are primarily involved in nonvesicular transport of lipids between membranes (14Alpy F. Tomasetto C. START ships lipids across interorganelle space.Biochimie. 2014; 96: 85-95Crossref PubMed Scopus (59) Google Scholar). For instance, STARD4 is a START domain protein that binds and transports cholesterol from the plasma membrane to mitochondria, the ER, and the endocytic recycling compartment, equilibrating cholesterol content among cellular membranes to fit their biophysical properties and physiological needs (15Martin L.A. Kennedy B.E. Karten B. Mitochondrial cholesterol: mechanisms of import and effects on mitochondrial function.J. Bioenerg. Biomembr. 2016; 48: 137-151Crossref PubMed Scopus (34) Google Scholar). Purified CC1736 binds to CoQn with variable polyisoprenoid chain lengths and to the farnesylated analog of a late-stage CoQ intermediate, demethoxy-CoQ3 (DMQ3) (9Allan C.M. Hill S. Morvaridi S. Saiki R. Johnson J.S. Liau W.S. Hirano K. Kawashima T. Ji Z. Loo J.A. et al.A conserved START domain coenzyme Q-binding polypeptide is required for efficient Q biosynthesis, respiratory electron transport, and antioxidant function in Saccharomyces cerevisiae.Biochim. Biophys. Acta. 2013; 1831: 776-791Crossref PubMed Scopus (23) Google Scholar). Coq10 polypeptides isolated from S. cerevisiae and S. pombe copurify with CoQ6 and CoQ10, respectively (10Barros M.H. Johnson A. Gin P. Marbois B.N. Clarke C.F. Tzagoloff A. The Saccharomyces cerevisiae COQ10 gene encodes a START domain protein required for function of coenzyme Q in respiration.J. Biol. Chem. 2005; 280: 42627-42635Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar, 16Cui T.Z. Kawamukai M. Coq10, a mitochondrial coenzyme Q binding protein, is required for proper respiration in Schizosaccharomyces pombe.FEBS J. 2009; 276: 748-759Crossref PubMed Scopus (30) Google Scholar). Similarly, CoQ8 copurifies with the S. pombe Coq10 polypeptide expressed in E. coli (16Cui T.Z. Kawamukai M. Coq10, a mitochondrial coenzyme Q binding protein, is required for proper respiration in Schizosaccharomyces pombe.FEBS J. 2009; 276: 748-759Crossref PubMed Scopus (30) Google Scholar). These observations have led to the current hypothesis that the Coq10 polypeptide is a putative CoQn chaperone, necessary for delivering CoQ from its site of synthesis and/or the pool of free CoQ to sites of function. Complex III inhibitors, antimycin A and myxothiazol, enhance reactive oxygen species (ROS) formation by blocking oxidation of cytochrome bH at the N-site or inhibiting reduction of cytochrome bL at the P-site, respectively (17Wikström M.K. Berden J.A. Oxidoreduction of cytochrome b in the presence of antimycin.Biochim. Biophys. Acta. 1972; 283: 403-420Crossref PubMed Scopus (201) Google Scholar, 18Starkov A.A. Fiskum G. Myxothiazol induces H2O2 production from mitochondrial respiratory chain.Biochem. Biophys. Res. Commun. 2001; 281: 645-650Crossref PubMed Scopus (105) Google Scholar, 19Dröse S. Brandt U. The mechanism of mitochondrial superoxide production by the cytochrome bc1 complex.J. Biol. Chem. 2008; 283: 21649-21654Abstract Full Text Full Text PDF PubMed Scopus (252) Google Scholar). Thereby, antimycin A induces ROS through reverse electron flow from cytochrome bL to CoQ to form the semiquinone radical (20Busso C. Tahara E.B. Ogusucu R. Augusto O. Ferreira-Junior J.R. Tzagoloff A. Kowaltowski A.J. Barros M.H. Saccharomyces cerevisiae coq10 null mutants are responsive to antimycin A.FEBS J. 2010; 277: 4530-4538Crossref PubMed Scopus (16) Google Scholar), whereas myxothiazol-dependent ROS production results from incomplete CoQH2 oxidation by slow reduction of the Rieske iron-sulfur protein (18Starkov A.A. Fiskum G. Myxothiazol induces H2O2 production from mitochondrial respiratory chain.Biochem. Biophys. Res. Commun. 2001; 281: 645-650Crossref PubMed Scopus (105) Google Scholar, 19Dröse S. Brandt U. The mechanism of mitochondrial superoxide production by the cytochrome bc1 complex.J. Biol. Chem. 2008; 283: 21649-21654Abstract Full Text Full Text PDF PubMed Scopus (252) Google Scholar). Mitochondria isolated from yeast coq10Δ produce significantly elevated ROS in the presence of antimycin A, but not myxothiazol, suggesting that in the absence of the Coq10 polypeptide, electron transfer from CoQH2 to the Rieske iron-sulfur protein is defective (20Busso C. Tahara E.B. Ogusucu R. Augusto O. Ferreira-Junior J.R. Tzagoloff A. Kowaltowski A.J. Barros M.H. Saccharomyces cerevisiae coq10 null mutants are responsive to antimycin A.FEBS J. 2010; 277: 4530-4538Crossref PubMed Scopus (16) Google Scholar). This specific requirement for the presence of the Coq10 START domain polypeptide for functional electron transfer by complex III is further substantiated by the binding of both oxidized and reduced forms of a photo-reactive azido-quinone probe to the Coq10 polypeptide (21Murai M. Matsunobu K. Kudo S. Ifuku K. Kawamukai M. Miyoshi H. Identification of the binding site of the quinone-head group in mitochondrial Coq10 by photoaffinity labeling.Biochemistry. 2014; 53: 3995-4003Crossref PubMed Scopus (10) Google Scholar). CoQ deficiencies are associated with human disease and the beneficial effects of CoQ10 supplementation in therapeutic regimens are increasingly appreciated (1Alcázar-Fabra M. Trevisson E. Brea-Calvo G. Clinical syndromes associated with coenzyme Q10 deficiency.Essays Biochem. 2018; 62: 377-398Crossref PubMed Scopus (32) Google Scholar, 4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar). Mutations in several genes encoding CoQ biosynthetic enzymes result in primary CoQ deficiency and cause encephalopathy, cerebellar ataxia, cardiomyopathy, nephrotic syndrome, and myopathy (1Alcázar-Fabra M. Trevisson E. Brea-Calvo G. Clinical syndromes associated with coenzyme Q10 deficiency.Essays Biochem. 2018; 62: 377-398Crossref PubMed Scopus (32) Google Scholar, 4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar). CoQ deficiency can also occur secondary to mutations in aprataxin, electron transfer flavoprotein dehydrogenase, or serine/threonine-protein kinase B-Raf (3Desbats M.A. Lunardi G. Doimo M. Trevisson E. Salviati L. Genetic bases and clinical manifestations of coenzyme Q10 (CoQ10) deficiency.J. Inherit. Metab. Dis. 2015; 38: 145-156Crossref PubMed Scopus (0) Google Scholar). CoQ10 supplementation rescues the proteinuria in patients with nephrotic syndrome, provided that therapy is initiated early (22Montini G. Malaventura C. Salviati L. Early coenzyme Q10 supplementation in primary coenzyme Q10 deficiency.N. Engl. J. Med. 2008; 358: 2849-2850Crossref PubMed Scopus (157) Google Scholar). Patients who develop myalgia under statin administration are often prompted to take CoQ10 supplements to mitigate adverse symptoms (23Marcoff L. Thompson P.D. The role of coenzyme Q10 in statin-associated myopathy: a systematic review.J. Am. Coll. Cardiol. 2007; 49: 2231-2237Crossref PubMed Scopus (386) Google Scholar). Long term CoQ10 treatment has also been shown to improve symptoms and reduce major adverse cardiovascular events when it is used as adjunctive treatment in patients with chronic heart failure (24Ayer A. Macdonald P. Stocker R. CoQ10 function and role in heart failure and ischemic heart disease.Annu. Rev. Nutr. 2015; 35: 175-213Crossref PubMed Scopus (26) Google Scholar, 25Mortensen S.A. Rosenfeldt F. Kumar A. Dolliner P. Filipiak K.J. Pella D. Alehagen U. Steurer G. Littarru G.P. Investigators Q.S.S. The effect of coenzyme Q10 on morbidity and mortality in chronic heart failure: results from Q-SYMBIO: a randomized double-blind trial.JACC Heart Fail. 2014; 2: 641-649Crossref PubMed Scopus (199) Google Scholar). Yeast is a superb model organism in which to study CoQ biosynthesis because many of the enzymes involved in CoQ biosynthesis are functionally conserved from yeast to humans (4Stefely J.A. Pagliarini D.J. Biochemistry of mitochondrial coenzyme Q biosynthesis.Trends Biochem. Sci. 2017; 42: 824-843Abstract Full Text Full Text PDF PubMed Scopus (73) Google Scholar, 5Awad A.M. Bradley M.C. Fernández-del-Río L. Nag A. Tsui H.S. Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans.Essays Biochem. 2018; 62: 361-376Crossref PubMed Scopus (40) Google Scholar). In this work, we test the human co-orthologs of yeast Coq10, COQ10A and COQ10B, for their ability to complement the yeast coq10Δ mutant. We show that expression of human COQ10A or COQ10B rescues yeast coq10Δ-defective respiration and its sensitivity to oxidative stress, and restores steady-state levels of Coq polypeptides. However, neither COQ10A nor COQ10B expression is able to stabilize the yeast CoQ synthome or rescue the partial defect in de novo CoQ6 biosynthesis characteristic of the yeast coq10Δ mutant. S. cerevisiae strains used in this study are described in Table 1. Growth media for yeast included YPD (1% Bacto yeast extract, 2% Bacto peptone, 2% dextrose), YPG (1% Bacto yeast extract, 2% Bacto peptone, 3% glycerol), and YPGal (1% Bacto yeast extract, 2% Bacto peptone, 2% galactose, 0.1% dextrose) (26Burke, D., D., Dawson, and T., Stearns, . 2000. Methods in Yeast Genetics. Cold Spring Harbor Laboratory Press, Plainview, NY.Google Scholar). Synthetic dextrose/minimal-complete (SD-Complete) and synthetic dextrose/minimal minus uracil (SD-Ura) [0.18% Difco yeast nitrogen base without amino acids and ammonium sulfate, 0.5% (NH4)2SO4, 0.14% NaH2PO4, 2% dextrose, complete amino acid supplement, or amino acid supplement lacking uracil] were prepared as described (27Barkovich R.J. Shtanko A. Shepherd J.A. Lee P.T. Myles D.C. Tzagoloff A. Clarke C.F. Characterization of the COQ5 gene from Saccharomyces cerevisiae. Evidence for a C-methyltransferase in ubiquinone biosynthesis.J. Biol. Chem. 1997; 272: 9182-9188Abstract Full Text Full Text PDF PubMed Scopus (71) Google Scholar). Solid media contained an additional 2% Bacto agar.TABLE 1Genotype and source of yeast strainsStrainGenotypeaMating type a (MAT a) is in bold to distinguish it from mating type α (MAT α).SourceW303 1BMAT α, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1R. RothsteinbDr. Rodney Rothstein, Department of Human Genetics, Columbia University, New York, NY.W303 coq1ΔMAT α, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq1::LEU2(99Gin P. Clarke C.F. Genetic evidence for a multi-subunit complex in coenzyme Q biosynthesis in yeast and the role of the Coq1 hexaprenyl diphosphate synthase.J. Biol. Chem. 2005; 280: 2676-2681Abstract Full Text Full Text PDF PubMed Scopus (64) Google Scholar)CC303MAT α, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq3::LEU2(100Do T.Q. Schultz J.R. Clarke C.F. Enhanced sensitivity of ubiquinone-deficient mutants of Saccharomyces cerevisiae to products of autoxidized polyunsaturated fatty acids.Proc. Natl. Acad. Sci. USA. 1996; 93: 7534-7539Crossref PubMed Scopus (0) Google Scholar)W303 coq4ΔMAT a, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq4::TRP1(101Hsu A.Y. Do T.Q. Lee P.T. Clarke C.F. Genetic evidence for a multi-subunit complex in the O-methyltransferase steps of coenzyme Q biosynthesis.Biochim. Biophys. Acta. 2000; 1484: 287-297Crossref PubMed Scopus (0) Google Scholar)W303 coq5ΔMAT α, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq5::HIS3(27Barkovich R.J. Shtanko A. Shepherd J.A. Lee P.T. Myles D.C. Tzagoloff A. Clarke C.F. Characterization of the COQ5 gene from Saccharomyces cerevisiae. Evidence for a C-methyltransferase in ubiquinone biosynthesis.J. Biol. Chem. 1997; 272: 9182-9188Abstract Full Text Full Text PDF PubMed Scopus (71) Google Scholar)W303 coq6ΔMAT a, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq6::LEU2(102Gin P. Hsu A.Y. Rothman S.C. Jonassen T. Lee P.T. Tzagoloff A. Clarke C.F. The Saccharomyces cerevisiae COQ6 gene encodes a mitochondrial flavin-dependent monooxygenase required for coenzyme Q biosynthesis.J. Biol. Chem. 2003; 278: 25308-25316Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar)W303 coq7ΔMAT α, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq7::LEU2(103Marbois B.N. Clarke C.F. The COQ7 gene encodes a protein in Saccharomyces cerevisiae necessary for ubiquinone biosynthesis.J. Biol. Chem. 1996; 271: 2995-3004Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar)W303 coq8ΔMAT a, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq8::HIS3(101Hsu A.Y. Do T.Q. Lee P.T. Clarke C.F. Genetic evidence for a multi-subunit complex in the O-methyltransferase steps of coenzyme Q biosynthesis.Biochim. Biophys. Acta. 2000; 1484: 287-297Crossref PubMed Scopus (0) Google Scholar)W303 coq9ΔMAT α, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq9::URA3(104Johnson A. Gin P. Marbois B.N. Hsieh E.J. Wu M. Barros M.H. Clarke C.F. Tzagoloff A. COQ9, a new gene required for the biosynthesis of coenzyme Q in Saccharomyces cerevisiae.J. Biol. Chem. 2005; 280: 31397-31404Abstract Full Text Full Text PDF PubMed Scopus (72) Google Scholar)W303 coq10ΔMAT a, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq10::HIS3(10Barros M.H. Johnson A. Gin P. Marbois B.N. Clarke C.F. Tzagoloff A. The Saccharomyces cerevisiae COQ10 gene encodes a START domain protein required for function of coenzyme Q in respiration.J. Biol. Chem. 2005; 280: 42627-42635Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar)W303 coq11ΔMAT a, ade2-1 can1-100 his3-11,15 leu2-3,112 trp1-1 ura3-1 coq11::HIS3This studya Mating type a (MAT a) is in bold to distinguish it from mating type α (MAT α).b Dr. Rodney Rothstein, Department of Human Genetics, Columbia University, New York, NY. Open table in a new tab Plasmids used in this study are listed in Table 2. Generation of single-copy (pQM) and multi-copy (pRCM) yeast expression vectors was previously described (9Allan C.M. Hill S. Morvaridi S. Saiki R. Johnson J.S. Liau W.S. Hirano K. Kawashima T. Ji Z. Loo J.A. et al.A conserved START domain coenzyme Q-binding polypeptide is required for efficient Q biosynthesis, respiratory electron transport, and antioxidant function in Saccharomyces cerevisiae.Biochim. Biophys. Acta. 2013; 1831: 776-791Crossref PubMed Scopus (23) Google Scholar, 28Hsu A.Y. Poon W.W. Shepherd J.A. Myles D.C. Clarke C.F. Complementation of coq3 mutant yeast by mitochondrial targeting of the Escherichia coli UbiG polypeptide: evidence that UbiG catalyzes both O-methylation steps in ubiquinone biosynthesis.Biochemistry. 1996; 35: 9797-9806Crossref PubMed Scopus (91) Google Scholar). Both pQM and pRCM contain the yeast CYC1 promoter and the first 35 residues of the yeast COQ3 ORF, corresponding to the proposed Coq3 mitochondrial leader sequence to direct import of human proteins into yeast mitochondria. To generate the single- and multi-copy yeast expression vectors of human COQ10A, the human COQ10A ORF (mRNA #1, Fig. 1A), encoding residues 44-247, was PCR amplified from pHCOQ10/ST1 (10Barros M.H. Johnson A. Gin P. Marbois B.N. Clarke C.F. Tzagoloff A. The Saccharomyces cerevisiae COQ10 gene encodes a START domain protein required for function of coenzyme Q in respiration.J. Biol. Chem. 2005; 280: 42627-42635Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar) with primers 5′-ggccATCGATATGAGGTTTCTGACCTCCTGC-3′ and 5′-ggccGGTACCTCAAGTCTGGTGCACCTC-3′, and cloned into pQM and pRCM vectors using the restriction enzymes, ClaI and KpnI (New England BioLabs), to generate pQM COQ10A and pRCM COQ10A, respectively. Similarly, full-length human COQ10B ORF (mRNA #1, Fig. 1A), encoding residues 1-238, was PCR amplified from COQ10B cDNA clone (GeneCopoeia) with primers 5′-ggccATCGATATGGCAGCTCGGACTGGTCAT-3′ and 5′-ggccGGTACCTTATGTGTGATGGACTTCATGAAGCATTAACTCC-3′ to generate pQM COQ10B and pRCM COQ10B.TABLE 2Yeast expression vectorsPlasmidRelevant Genes/MarkersSourcepQMpAH01 with COQ3 mito leader, single-copy(28Hsu A.Y. Poon W.W. Shepherd J.A. Myles D.C. Clarke C.F. Complementation of coq3 mutant yeast by mitochondrial targeting of the Escherichia coli UbiG polypeptide: evidence that UbiG catalyzes both O-methylation steps in ubiquinone biosynthesis.Biochemistry. 1996; 35: 9797-9806Crossref PubMed Scopus (91) Google Scholar)pQM COQ10ApQM with human COQ10A; single-copyThis workpQM COQ10BpQM with human COQ10B; single-copyThis workpRCMpCH1 with COQ3 mito leader; multi-copy(9Allan C.M. Hill S. Morvaridi S. Saiki R. Johnson J.S. Liau W.S. Hirano K. Kawashima T. Ji Z. Loo J.A. et al.A conserved START domain coenzyme Q-binding polypeptide is required for efficient Q biosynthesis, respiratory electron transport, and antioxidant function in Saccharomyces cerevisiae.Biochim. Biophys. Acta. 2013; 1831: 776-791Crossref PubMed Scopus (23) Google Scholar)pRCM COQ10ApRCM with human COQ10A; multi-copyThis workpRCM COQ10BpRCM with human COQ10B; multi-copyThis work Open table in a new tab Each of the following plasmids, pQM (empty vector), pQM C" @default.
• W2943082710 created "2019-05-09" @default.
• W2943082710 creator A5001167526 @default.
• W2943082710 creator A5007715260 @default.
• W2943082710 creator A5013520775 @default.
• W2943082710 creator A5026118622 @default.
• W2943082710 creator A5053012801 @default.
• W2943082710 creator A5053230113 @default.
• W2943082710 creator A5056444335 @default.
• W2943082710 creator A5058254000 @default.
• W2943082710 creator A5065053978 @default.
• W2943082710 creator A5081322844 @default.
• W2943082710 date "2019-07-01" @default.
• W2943082710 modified "2023-10-06" @default.
• W2943082710 title "Human COQ10A and COQ10B are distinct lipid-binding START domain proteins required for coenzyme Q function" @default.
• W2943082710 cites W1193855731 @default.
• W2943082710 cites W1527231409 @default.
• W2943082710 cites W1555471479 @default.
• W2943082710 cites W1588290914 @default.
• W2943082710 cites W1647674345 @default.
• W2943082710 cites W1657534147 @default.
• W2943082710 cites W1701273068 @default.
• W2943082710 cites W1823885789 @default.
• W2943082710 cites W1860616385 @default.
• W2943082710 cites W1963530308 @default.
• W2943082710 cites W1963587806 @default.
• W2943082710 cites W1963917155 @default.
• W2943082710 cites W1965634593 @default.
• W2943082710 cites W1967433085 @default.
• W2943082710 cites W1969102813 @default.
• W2943082710 cites W1970891030 @default.
• W2943082710 cites W1971668538 @default.
• W2943082710 cites W1971714188 @default.
• W2943082710 cites W1975778017 @default.
• W2943082710 cites W1977811408 @default.
• W2943082710 cites W1979088319 @default.
• W2943082710 cites W1982085892 @default.
• W2943082710 cites W1987078348 @default.
• W2943082710 cites W1987102030 @default.
• W2943082710 cites W1987370810 @default.
• W2943082710 cites W1987513243 @default.
• W2943082710 cites W1987515388 @default.
• W2943082710 cites W1990249966 @default.
• W2943082710 cites W1993948951 @default.
• W2943082710 cites W1997338312 @default.
• W2943082710 cites W1997527771 @default.
• W2943082710 cites W2004377868 @default.
• W2943082710 cites W2005650337 @default.
• W2943082710 cites W2008524932 @default.
• W2943082710 cites W2012034410 @default.
• W2943082710 cites W2022482177 @default.
• W2943082710 cites W2023627793 @default.
• W2943082710 cites W2024102643 @default.
• W2943082710 cites W2028302194 @default.
• W2943082710 cites W2036512300 @default.
• W2943082710 cites W2037809759 @default.
• W2943082710 cites W2038076808 @default.
• W2943082710 cites W2042450476 @default.
• W2943082710 cites W2048971937 @default.
• W2943082710 cites W2056721274 @default.
• W2943082710 cites W2060035882 @default.
• W2943082710 cites W2061764676 @default.
• W2943082710 cites W2068119233 @default.
• W2943082710 cites W2070838021 @default.
• W2943082710 cites W2071486470 @default.
• W2943082710 cites W2074189512 @default.
• W2943082710 cites W2075184035 @default.
• W2943082710 cites W2076135395 @default.
• W2943082710 cites W2085058795 @default.
• W2943082710 cites W2089752219 @default.
• W2943082710 cites W2092143550 @default.
• W2943082710 cites W2095389529 @default.
• W2943082710 cites W2096704460 @default.
• W2943082710 cites W2102993309 @default.
• W2943082710 cites W2103096171 @default.
• W2943082710 cites W2111647009 @default.
• W2943082710 cites W2112464428 @default.
• W2943082710 cites W2118203012 @default.
• W2943082710 cites W2118868063 @default.
• W2943082710 cites W2119677134 @default.
• W2943082710 cites W2120230374 @default.
• W2943082710 cites W2125842439 @default.
• W2943082710 cites W2129639464 @default.
• W2943082710 cites W2134499258 @default.
• W2943082710 cites W2136466059 @default.
• W2943082710 cites W2140488719 @default.
• W2943082710 cites W2141026950 @default.
• W2943082710 cites W2143965041 @default.
• W2943082710 cites W2146481406 @default.
• W2943082710 cites W2151581834 @default.
• W2943082710 cites W2154714625 @default.
• W2943082710 cites W2156235607 @default.
• W2943082710 cites W2156654965 @default.
• W2943082710 cites W2157369531 @default.
• W2943082710 cites W2159675211 @default.
• W2943082710 cites W2160378127 @default.
• W2943082710 cites W2162980545 @default.
• W2943082710 cites W2167345470 @default.

• next