CYTOCHROME C OXIDASE IN HEALTH AND DISEASE

描述（由申请人提供）：本提案是对资助公告NOT-OD-09-058的回应，标题为：NIH宣布恢复法案资金的可用性，用于竞争性修订申请，以请求为我的资助5 R 01 GM 071775 -04提供竞争性补充; Barrientos，Antoni（PI）02/01/06-1/31/11，题为“健康和疾病中的细胞色素C氧化酶”。 母公司和拟议的研究的主要目的是调查线粒体细胞色素c氧化酶（考克斯）在野生型细胞和进化保守的考克斯组装因子突变的细胞中的生物起源。考克斯缺乏是人类线粒体脑肌病的最常见原因。更好地了解考克斯生物起源是阐明这组疾病的分子基础是必不可少的。 本研究以酿酒酵母为研究模型，探讨考克斯的生物合成。我们已经获得了翻译调控系统的证据，通过该系统，可调控的编码考克斯亚基1的合成由其装配伙伴的可用性，从而起搏Cox 1 p的合成，以利用其在装配过程中。已经鉴定了几种蛋白质参与该调节系统，包括翻译激活剂Mss 51 p和考克斯组装因子Cox 14 p和Shy 1 p。Ms 51 p与COX 1 mRNA相互作用以促进翻译，并与新合成的Cox 1 p相互作用以促进其成熟，形成由Cox 14 p稳定的复合物。当Cox 1 p通过Shy 1 p催化的一个步骤在组装过程的下游进行时，复合物被破坏，而Mss 51 p则被回收到新的一轮翻译中，正如在母基金中提出的那样。有趣的是，Mss 51 p的过表达恢复了shy 1突变体的考克斯组装和呼吸能力。 考克斯装配是显着保守的从酵母到人。SURF 1是Shy 1 p的人类同源物，已被充分表征，并已发现在患有与考克斯缺陷相关的线粒体脑肌病的患者中发生突变。相反，翻译激活剂Mss 51 p的人类对应物尚未被鉴定。通过BLAST分析，我们现在已经认识到人类基因ZMYND 17，位于染色体10作为一个遥远的Mss 51 p同源。在本申请中，我们建议使用人细胞培养物来表征ZMYND 17、其功能及其在SURF 1缺陷的人成纤维细胞中恢复考克斯生物发生的能力。 公共卫生相关性：该提案是对资助公告NOT-OD-09-058的回应，标题为：NIH宣布恢复法案资金的可用性，用于竞争性修订申请，以请求为我的赠款5 R 01 GM 071775 -04提供竞争性补充; Barrientos，Antoni（PI）02/01/06-1/31/11，题为“健康和疾病中的细胞色素C氧化酶”。该建议的主要焦点是对人基因ZMYND 17的产物的表征，ZMYND 17是酵母COX 1 mRNA特异性翻译因子Mss 51 p的推定同源物。在本申请中，我们提出使用人细胞培养物来表征ZMYND 17及其功能。

DESCRIPTION (provided by applicant): This proposal is a response to the funding announcement NOT-OD-09-058 with title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications to request for a Competitive Supplement for my grant 5R01GM071775-04; Barrientos, Antoni (PI) 02/01/06-1/31/11 entitled "CYTOCHROME C OXIDASE IN HEALTH AND DISEASE". The main objective of the parent and proposed research is to investigate the biogenesis of mitochondrial cytochrome c oxidase (COX) in wild type cells and in cells with mutations in evolutionary conserved COX assembly factors. COX deficiency is the most frequent cause of mitochondrial encephalomyopathies in humans. A better understanding of COX biogenesis is essential for elucidating the molecular basis underlying this group of diseases. Following the project proposed in the parent grant, we are using the yeast Saccharomyces cerevisiae as a research model to study COX biogenesis. We have obtained evidence of a translational regulatory system by which the synthesis of the mitochondrially encoded COX subunit 1 is regulated by the availability of its assembly partners thereby pacing Cox1p synthesis to its utilization during assembly. Several proteins have been identified to be involved in this regulatory system, including the translational activator Mss51p and the COX assembly factors Cox14p and Shy1p. Ms51p interacts with both the COX1 mRNA to promote translation and with the newly synthesized Cox1p, possibly to facilitate its maturation, forming a complex that is stabilizazed by Cox14p. The complex is disrupted when Cox1p proceeds downstream the assembly process by a step catalyzed by Shy1p and Mss51p is recycled to new rounds of translation as proposed in the parent grant. Interestingly, overexpression of Mss51p restores COX assembly and respiratory competence to mutants of shy1. COX assembly is signficantly conserved from yeast to human. SURF1, the human homologue of Shy1p is well characterized and has been found mutated in patients suffering from mitochondrial encephalomyopathies associated with COX deficiency. Instead, the human counterpart of the translational activator Mss51p has not been yet identified. By BLAST analyses we have now recognized the human gene ZMYND17, located in chromosome 10 as a distant Mss51p homologue. In this application we propose to use human cell cultures to characterize ZMYND17, its function and its ability to restoring COX biogenesis in SURF1-deficient human fibroblasts. PUBLIC HEALTH RELEVANCE: This proposal is a response to the funding announcement NOT-OD-09-058 with title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications to request for a Competitive Supplement for my grant 5R01GM071775-04; Barrientos, Antoni (PI) 02/01/06-1/31/11 entitled "CYTOCHROME C OXIDASE IN HEALTH AND DISEASE". The main focus of this proposal is on the characterization the product of the human gene ZMYND17, a putative homologue of the yeast COX1 mRNA - specific translational factor Mss51p. In this application we propose to use human cell cultures to characterize ZMYND17 and its function.