Amino-terminal acetylation of proteins in mammalian biology and disease

哺乳动物生物学和疾病中蛋白质的氨基末端乙酰化

• 批准号: 10604927
• 负责人: GHOLSON LYON
• 金额: $ 23.86万
• 依托单位: INSTITUTE FOR BASIC RES IN DEV DISABIL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10604927
• 关键词: Acetylation; Acetyltransferase; Adenosine Triphosphate; Affect; Alzheimer' s Disease; Binding; Biology; Cardiac; Cardiomegaly; Cell Culture Techniques; Cell Death; Cell Line; Cells; Communities; Complex; Congenital Disorders; Congenital Heart Defects; Coupled; Craniofacial Abnormalities; Cryptorchidism; Development; Developmental Delay Disorders; Disease; Dissection; Energy Metabolism; Equipment; Family; Functional disorder; Genes; Genetic Diseases; Genus Hippocampus; Gold; Health; Heart failure; Homeostasis; Human; Huntington Disease; Laboratories; Link; Lysine; Malignant Neoplasms; Measurement; Metabolic; Missense Mutation; Mitochondria; Modification; Molecular Biology; Muscle hypotonia; Mutation; N-terminal; Names; Neurodegenerative Disorders; Parkinson Disease; Pathway interactions; Patients; Phenotype; Post-Translational Protein Processing; Production; Protein Acetylation; Proteins; Rare Diseases; Reporting; Role; Syndrome; Time; Utah; boys; congenital heart disorder; de novo mutation; in vivo Model; induced pluripotent stem cell; insight; instrument; knock-down; neurodevelopment; oxidation; protein function; tumor progression

PROJECT SUMMARY Although the scientific community has made substantial progress in elucidating the function of many genes, much remains unknown, particularly concerning the diversity introduced into proteins with co- and post- translational modifications. One such modification is amino- (or N-) terminal acetylation (NTA), which is considerably understudied, with very few reports on the mammalian N-terminome. Protein acetylation occurs both at lysine residues within proteins (lysine acetylation or N-ε-acetylation) and at the N-terminus of proteins (Nt-acetylation or N--acetylation). Protein Nt-acetylation is among the most common modifications of eukaryotic proteins and is carried out by N-terminal acetyltransferases (NATs). The knockdown phenotypes of human NATs in cell culture suggest that protein NTA is an essential modification in human cells to maintain proliferation, but functional insights and mammalian in vivo models are lacking. Understanding of a general role for NTA remains elusive, and only a few examples in which NTA affects protein function, complex formation, activity, or stability are known. My laboratory discovered and characterized the first genetic disease coupled to NTA of proteins, involving a missense mutation in the X-linked gene NAA10; we named this rare disease Ogden syndrome (OS) in honor of the hometown (Ogden, Utah), where the first family we identified with OS lived. The affected boys have a distinct combination of craniofacial anomalies, hypotonia, global developmental delays, cryptorchidism, cardiac anomalies, and cardiomegaly. We and others then found more than a dozen families with overlapping phenotypes with additional mutations in NAA10 in this pathway; we also reported recently that de novo mutations in NAA15, encoding a binding partner for NAA10, are involved in congenital heart defects and/or neurodevelopment. This finding is consistent with the range of cardiac anomalies and neurodevelopmental delays seen in OS (now more broadly known as NAA10-related disorders). As part of our focus on the mechanistic dissection of NTA, we are undertaking detailed metabolic characterization of patient- derived induced pluripotent stem cell lines. In this instrument supplement application, we are requesting purchase of an Agilent Seahorse machine, which is the gold standard for reliable, accurate, and routine measurements that can be applied to assess glycolytic rates, energy expenditure (e.g., cellular activation, proliferation, differentiation), adenosine triphosphate production rate in real-time, substrate oxidation, mitochondrial function, cell death, and general cellular homeostasis. This instrument will enable us to expand our study of the molecular biology and pathophysiology associated with NAA10- and NAA15-related disorders and the NTA pathway, as part of a sustained effort to understand the role of NTA in mammalian biology. These studies will be a critical step toward revealing the role of NTA in human health and disease, as NTA has been linked to cancer progression and neurodegenerative diseases, including Parkinson’s, Alzheimer’s, and Huntington’s diseases.

项目摘要 尽管科学界在阐明许多基因的功能方面取得了实质性进展， 还有很多未知的，特别是关于引入蛋白质的多样性， 翻译修饰一种这样的修饰是氨基-（或N-）末端乙酰化（NTA），其是 在哺乳动物的N-末端基因组中，研究还相当不足，只有很少的报道。蛋白质乙酰化发生 在蛋白质内的赖氨酸残基（赖氨酸乙酰化或N-ε-乙酰化）和蛋白质的N-末端 （N-乙酰化或N-乙酰化）。蛋白质N-乙酰化是最常见的修饰之一， 真核生物的蛋白质，并进行了N-末端乙酰转移酶（NAT）。基因敲低表型 细胞培养物中人NAT表明蛋白质NTA是人细胞中维持 增殖，但缺乏功能的见解和哺乳动物体内模型。对一般作用的理解 对于NTA仍然难以捉摸，并且只有少数NTA影响蛋白质功能，复合物形成， 活性或稳定性已知。我的实验室发现并描述了第一种遗传疾病， 蛋白质的NTA，涉及X连锁基因NAA 10的错义突变;我们将这种罕见疾病命名为Ogden 在家乡（奥格登，犹他州）的荣誉，在那里的第一个家庭，我们确定与OS生活。的 受影响的男孩具有颅面异常、张力减退、整体发育迟缓的明显组合， 隐睾、心脏畸形和心脏肥大。我们和其他人随后找到了十几个家庭 在这一途径中，NAA 10的表型与其他突变重叠;我们最近还报道， 编码NAA 10结合伴侣的NAA 15的新生突变与先天性心脏病有关 和/或神经发育。这一发现与心脏异常的范围一致， 在OS中观察到的神经发育延迟（现在更广泛地称为NAA 10相关疾病）。作为我们 专注于NTA的机制解剖，我们正在进行患者的详细代谢表征， 衍生的诱导多能干细胞系。在此仪器补充申请中，我们要求 购买Agilent Seahorse机器，这是可靠、准确和常规的黄金标准， 可以应用于评估糖酵解速率，能量消耗（例如，细胞活化， 增殖，分化），实时的三磷酸腺苷产生速率，底物氧化， 线粒体功能、细胞死亡和一般细胞内稳态。这一工具将使我们能够扩大 我们对NAA 10和NAA 15相关疾病的分子生物学和病理生理学的研究 和NTA途径，作为持续努力的一部分，以了解NTA在哺乳动物生物学中的作用。这些 研究将是揭示NTA在人类健康和疾病中作用的关键一步，因为NTA一直是 与癌症进展和神经退行性疾病有关，包括帕金森氏症，阿尔茨海默氏症， 亨廷顿氏病。