title: Tissue- and species-specific differences in cytochrome c oxidase assembly induced by SURF1 defects.
abstract: Mitochondrial protein SURF1 is a specific assembly factor of cytochrome c oxidase (COX), but its function is poorly understood. SURF1 gene mutations cause a severe COX deficiency manifesting as the Leigh syndrome in humans, whereas in mice SURF1(-/-) knockout leads only to a mild COX defect. We used SURF1(-/-) mouse model for detailed analysis of disturbed COX assembly and COX ability to incorporate into respiratory supercomplexes (SCs) in different tissues and fibroblasts. Furthermore, we compared fibroblasts from SURF1(-/-) mouse and SURF1 patients to reveal interspecies differences in kinetics of COX biogenesis using 2D electrophoresis, immunodetection, arrest of mitochondrial proteosynthesis and pulse-chase metabolic labeling. The crucial differences observed are an accumulation of abundant COX1 assembly intermediates, low content of COX monomer and preferential recruitment of COX into I-III2-IVn SCs in SURF1 patient fibroblasts, whereas SURF1(-/-) mouse fibroblasts were characterized by low content of COX1 assembly intermediates and milder decrease in COX monomer, which appeared more stable. This pattern was even less pronounced in SURF1(-/-) mouse liver and brain. Both the control and SURF1(-/-) mice revealed only negligible formation of the I-III2-IVn SCs and marked tissue differences in the contents of COX dimer and III2-IV SCs, also less noticeable in liver and brain than in heart and muscle. Our studies support the view that COX assembly is much more dependent on SURF1 in humans than in mice. We also demonstrate markedly lower ability of mouse COX to form I-III2-IVn supercomplexes, pointing to tissue-specific and species-specific differences in COX biogenesis.
authors.affiliation: Institute of Physiology of the Czech Academy of Sciences, Vídeňská 1083, Prague, Czech Republic.
authors.forename: Nikola
authors.initials: N
authors.lastname: Kovářová
authors.affiliation: Institute of Physiology of the Czech Academy of Sciences, Vídeňská 1083, Prague, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Pecina
authors.affiliation: Institute of Physiology of the Czech Academy of Sciences, Vídeňská 1083, Prague, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Nůsková
authors.affiliation: Institute of Physiology of the Czech Academy of Sciences, Vídeňská 1083, Prague, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Vrbacký
authors.affiliation: Molecular Neurogenetics Unit, Instituto Neurologico "C. Besta", via Temolo 4, 20126 Milan, Italy; MRC-Mitochondrial Biology Unit, Wellcome Trust MRC Bldg, Addenbrookes Hospital Hills Rd, Cambridge CB2 0XY, UK.
authors.forename: Massimo
authors.initials: M
authors.lastname: Zeviani
authors.affiliation: Institute of Physiology of the Czech Academy of Sciences, Vídeňská 1083, Prague, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Mráček
authors.affiliation: MRC-Mitochondrial Biology Unit, Wellcome Trust MRC Bldg, Addenbrookes Hospital Hills Rd, Cambridge CB2 0XY, UK.
authors.forename: Carlo
authors.initials: C
authors.lastname: Viscomi
authors.affiliation: Institute of Physiology of the Czech Academy of Sciences, Vídeňská 1083, Prague, Czech Republic. Electronic address: houstek@biomed.cas.cz.
authors.forename: Josef
authors.initials: J
authors.lastname: Houštěk
mesh_terms: D000818:Animals; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007888:Leigh Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D009928:Organ Specificity; D013045:Species Specificity
chemical_list: D008565:Membrane Proteins; D024101:Mitochondrial Proteins; C092928:Surf-1 protein; D003576:Electron Transport Complex IV
keywords: Cytochrome c oxidase; Doxycycline; Leigh syndrome; Pulse-chase; Respiratory supercomplexes; SURF1(−/−) mouse knockout