Decreased amounts of core proteins I and II and the iron-sulfur protein in mitochondria from yeast lacking cytochrome b but containing cytochrome c1 |
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| Authors: | K Sen D S Beattie |
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| Affiliation: | 1. Institute of Forensic Medicine, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Albertstrasse 9, Freiburg 79104, Germany;2. Institute of Legal Medicine, Medical University of Innsbruck, Muellerstrasse 44, Innsbruck 6020, Austria;3. i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal;4. Forensic Science Program, The Pennsylvania State University, USA;1. Pediatric Haematology Oncology Unit, Department of Pediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh, India;4. Pediatric Biochemistry Laboratory, Department of Pediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh, India;2. Department of Haematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India;3. Department of Clinical Haematology and Medical Oncology, Postgraduate Institute of Medical Education and Research, Chandigarh, India;5. Department of Pathology, Boston Children''s Hospital, Boston, Massachusetts;1. Division of Genetics and Epidemiology, The Institute of Cancer Research, Sutton, United Kingdom;2. Sheffield Diagnostic Genetics Service, NHS North East and Yorkshire Genomic Laboratory Hub, Sheffield Children''s NHS Foundation Trust, Sheffield, United Kingdom;3. Wessex Regional Genetics Laboratory, Central and South Genomics Laboratory Hub, Salisbury NHS Foundation Trust, Salisbury District Hospital, Salisbury, Wiltshire, United Kingdom;4. Cambridge Genomic Laboratory, East Genomic Laboratory Hub, Cambridge University Hospital NHS Foundation Trust, Cambridge, United Kingdom;5. Manchester Centre for Genomic Medicine and North West Genomic Laboratory Hub, Manchester University NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, United Kingdom;6. North East and Yorkshire Genomic Laboratory Hub, The Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom;7. Bristol Genetics Laboratory, Southmead Hospital, North Bristol NHS Trust, Bristol, United Kingdom;8. Cancer Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom;9. Human Genetics and Genomic Medicine, Faculty of Medicine, University of Southampton, Southampton, United Kingdom;10. Exeter Genomics Laboratory, South West Genomic Laboratory Hub, Royal Devon and Exeter NHS Foundation Trust, Exeter, United Kingdom;11. University of Exeter Medical School, Exeter, United Kingdom;12. Genomics England, London, United Kingdom;13. Division of Evolution & Genomic Sciences, The University of Manchester, Manchester, United Kingdom;14. Southeast Thames Regional Genetics Service, Guy''s and St Thomas'' NHS Foundation Trust, London, United Kingdom;15. Department of Medical Genetics, National Institute for Health Research Cambridge Biomedical Research Centre, University of Cambridge, Cambridge, United Kingdom;16. Wellcome Centre for Human Genetics/Centre for Personalised Medicine, University of Oxford, Oxford, United Kingdom;17. Clinical Ethics and Law, Faculty of Medicine, University of Southampton, Southampton, United Kingdom;18. Department of Clinical Genetics, St. George''s University Hospitals NHS Foundation Trust, London, United Kingdom;19. Cancer Genetics Unit, The Royal Marsden NHS Foundation Trust, London, United Kingdom;1. Division of Medical Genetics and Genomics, and Department of Genetic and Metabolic Diseases, The Children''s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, Zhejiang 310052, China;2. Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China;3. Department of Otorhinolaryngology-Head and Neck Surgery, Shandong Provincial ENT Hospital, Shandong University, Jinan, Shandong 250022, China;4. Department of Otorhinolaryngology Head and Neck Surgery, The Children''s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, Zhejiang 310052, China;1. Unitat d’Antropologia Biològica, Departament BAVE, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Barcelona, Spain;2. Department of Genetics and Microbiology, Faculty of Biosciences, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Barcelona, Spain;3. Institut de Biotecnologia i Biomedicina; Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Barcelona, Spain;4. Department of Pathology, Germans Trias i Pujol Research Institute, Badalona 08916 Barcelona, Spain;5. Dirección General de Salud Pública. Consejería de Sanidad. Junta de Castilla y León, 47007 Valladolid, Spain |
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| Abstract: | The effect of cytochrome b on the assembly of the subunits of complex III into the inner mitochondrial membrane has been studied in four mutants of yeast that lack a spectrally detectable cytochrome b and do not synthesize apocytochrome b. Quantitative analysis of intact mitochondria by immunoprecipitation or immunoblotting techniques with specific antisera revealed that the core proteins and the iron-sulfur protein were decreased 50% or more in the mitochondria from the mutants as compared to the wild type. Sonication of wild-type mitochondria did not result in any decrease in any of these proteins from the membrane; however, sonication of mitochondria from the four mutants resulted in a further decrease in the amount of these proteins suggesting that they are not as tightly bound to the mitochondrial membrane in the absence of cytochrome b. By contrast, the amounts of cytochrome c1 in the mitochondria, as determined both spectroscopically and immunologically, were not significantly affected by the absence of cytochrome b. In addition, no loss of cytochrome c1 was observed after sonication of the mitochondria suggesting that this protein is tightly bound to the membrane. These results suggest that the processing and/or assembly of these subunits of complex III into the mitochondrial membrane is affected by the absence of cytochrome b. |
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