Cytochrome Oxidase Polymorphisms in Aging

衰老中的细胞色素氧化酶多态性

• 批准号: 7228058
• 负责人: RUSSELL H. SWERDLOW
• 金额: $ 6.48万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-15 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7228058
• 关键词: ATP Synthesis Pathway; Address; Age; Aging; Amino Acids; Antioxidants; Base Sequence; Biochemical; Cell Line; Cells; Complementary DNA; Complex; Data; Electron Transport; Event; Fluorescent Probes; General Population; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genome; Glioma; Hereditary Disease; Human; Individual; Knock-out; Measurement; Mitochondria; Mitochondrial DNA; Multienzyme Complexes; Nervous system structure; Neurodegenerative Disorders; Nuclear; Oxygen; Play; Population; Production; Protein Subunits; Proteins; Reactive Oxygen Species; Research Personnel; Role; Standards of Weights and Measures; Techniques; Teratocarcinoma; Transfection; U251; Virginia; Work; base; cytochrome c oxidase; enzyme activity; homologous recombination; knockout gene; mitochondrial dysfunction; osteosarcoma; programs; restoration; rho; senescence

DESCRIPTION (provided by applicant): Some suspect mitochondria play a role in nervous system senescence and late-onset neurodegenerative disease. Data suggest relevant mechanisms may include inadequate electron transport chain (ETC) function and/or ETC-derived reactive oxygen species (ROS) production. The underlying basis for these ETC-associated events is, however, unknown. The ETC/ATP synthesis apparatus consists of five multimeric protein complexes. Four of them derive subunits from both mitochondrial and nuclear genes. Amino acid-changing polymorphisms are known for many of these genes. This proposal will determine whether ETC gene polymorphisms influence ETC activity and ROS production. It is hypothesized particular nuclear-mitochondrial ETC gene polymorphism combinations produce "efficient" enzyme complexes with robust activity, while other combinations produce "inefficient" enzyme complexes that are less kinetically robust. It is further hypothesized inefficient ETC enzyme complexes will produce more ROS than efficient enzyme complexes. To address this hypothesis, we will (1) determine nucleotide sequences of nuclear cytochrome oxidase (CO) genes from different mtDNA-depleted cell lines, as well as establish how common nuclear CO gene polymorphisms are in the general population; (2) identify how common mtDNA CO gene polymorphisms are in the general population; and (3) create a combination nuclear CO gene knockout-p0 cell line to facilitate replacement with particular mtDNA and nuclear CO gene variants. In humans, the mitochondrial ETC is the only place protein products from two distinct genomes directly interact. Because of genetic variability, multiple subunit combinations are possible. How mitochondrial DNA (mtDNA) and nuclear-derived ETC protein subunits mesh may determine whether an individual's nervous system ages successfully or not. This work could help explain why complex genetic diseases are sometimes associated with mitochondrial dysfunction.

描述（由申请人提供）：一些人怀疑线粒体在神经系统衰老和迟发性神经退行性疾病中发挥作用。数据表明相关机制可能包括电子传递链 (ETC) 功能不足和/或 ETC 衍生的活性氧 (ROS) 产生。然而，这些 ETC 相关事件的根本原因尚不清楚。 ETC/ATP 合成装置由五个多聚蛋白复合物组成。其中四个亚基来自线粒体和核基因。许多这些基因的氨基酸改变多态性是已知的。该提案将确定ETC基因多态性是否影响ETC活性和ROS产生。假设特定的核-线粒体ETC基因多态性组合产生具有强大活性的“高效”酶复合物，而其他组合产生动力学稳定性较差的“低效”酶复合物。进一步假设低效的 ETC 酶复合物将比高效的酶复合物产生更多的 ROS。为了解决这一假设，我们将（1）确定来自不同 mtDNA 耗尽细胞系的核细胞色素氧化酶（CO）基因的核苷酸序列，并确定核 CO 基因多态性在一般人群中的常见程度； (2) 确定线粒体DNA CO基因多态性在一般人群中的常见程度； (3) 创建组合核 CO 基因敲除 p0 细胞系，以促进特定 mtDNA 和核 CO 基因变体的替换。 在人类中，线粒体 ETC 是来自两个不同基因组的蛋白质产物直接相互作用的唯一场所。由于遗传变异性，多个亚基组合是可能的。线粒体 DNA (mtDNA) 和核衍生的 ETC 蛋白亚基如何啮合可能决定一个人的神经系统能否成功衰老。这项工作可以帮助解释为什么复杂的遗传疾病有时与线粒体功能障碍有关。