Cytochrome c oxidase subunit IV is essential for assembly and respiratory function of the enzyme complex.
Cytochrome c oxidase or complex IV, catalyzes the final step in mitochondrial electron transfer chain, and is regarded as one of the major regulation sites for oxidative phosphorylation. This enzyme is controlled by both nuclear and mitochondrial genomes. Among its 13 subunits, three are encoded by mitochondrial DNA and ten by nuclear DNA. In this work, an RNA interference approach was taken which led to the generation of mouse A9 cell derivatives with suppressed expression of nuclear-encoded subunit IV (COX IV) of this complex. The amounts of this subunit are decrease by 86% to 94% of normal level. A detail biosynthetic and functional analysis of several cell lines with suppressed COX IV expression revealed a loss of assembly of cytochrome c oxidase complex and, correspondingly, a reduction in cytochrome c oxidase-dependent respiration and total respiration. Furthermore, dysfunctional cytochrome c oxidase in the cells leads to a compromised mitochondrial membrane potential, a decreased ATP level, and failure to grow in galactose medium. Interestingly, suppression of COX IV expression also sensitizes the cells to apoptosis. These observations provide the evidence of the essential role of the COX IV subunit for a functional cytochrome c oxidase complex and also demonstrate a tight control of cytochrome c oxidase over oxidative phosphorylation. Finally, our results further shed some insights into the pathogenic mechanism of the diseases caused by dysfunctional cytochrome c oxidase complex.
Li
Youfen
Y
Department of Cellular and Structural Biology, University of Texas Health Sciences Center at San Antonio, 7703 Floyd Curl, San Antonio, Texas 78229, USA.
Park
Jeong-Soon
JS
Deng
Jian-Hong
JH
Bai
Yidong
Y
D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002460:Cell Line; D019069:Cell Respiration / Q000502:physiology; D003576:Electron Transport Complex IV / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D004586:Electrophoresis; D053078:Membrane Potential, Mitochondrial / Q000502:physiology; D051379:Mice; D009841:Oligonucleotides; D010085:Oxidative Phosphorylation; D010101:Oxygen Consumption / Q000502:physiology; D034622:RNA Interference
D009841:Oligonucleotides; D000255:Adenosine Triphosphate; D003576:Electron Transport Complex IV