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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概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，特别是关于通过共和和后引入蛋白质的多样性。一种这样的修饰是氨基（或N-）末端乙酰化（NTA），它是对此的研究相当少，关于哺乳动物 N 末端乙酰化发生的报道很少。位于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（Nt-乙酰化或 N-α-乙酰化）蛋白质 Nt-乙酰化是最常见的修饰之一。真核蛋白质并由 N 末端乙酰转移酶 (NAT) 进行敲低表型。细胞培养物中的人类 NAT 表明，蛋白质 NTA 是人类细胞维持正常状态的重要修饰。增殖，但缺乏对一般作用的功能见解和哺乳动物体内模型的了解。对于 NTA 仍然难以捉摸，只有少数例子表明 NTA 影响蛋白质功能、复合物形成、我的实验室发现并表征了第一种与此相关的遗传疾病。蛋白质 NTA，涉及 X 连锁基因 NAA10 的错义突变，我们将这种罕见疾病命名为 Ogden；综合症（OS），以纪念家乡（犹他州奥格登），我们发现的第一个患有 OS 的家庭就居住在那里。受影响的男孩具有明显的颅面异常、肌张力低下、整体发育迟缓的组合，我们和其他人随后发现了十多个家庭有隐睾症、心脏异常和心脏肥大。我们最近还报道了该通路中 NAA10 存在额外突变的重叠表型； NAA15 的从头突变编码 NAA10 的结合伴侣，与先天性心脏缺陷有关这一发现与心脏异常和/或神经发育的范围一致。 OS 中出现的神经发育迟缓（现在更广泛地称为 NAA10 相关疾病）。专注于 NTA 的机械解剖，我们正在对患者进行详细的代谢特征分析在此仪器补充申请中，我们要求获得衍生的诱导多能干细胞系。购买安捷伦 Seahorse 机器，这是可靠、准确和常规的黄金标准可用于评估糖酵解速率、能量消耗（例如细胞激活、增殖、分化）、实时三磷酸腺苷生产率、底物氧化、线粒体功能、细胞死亡和一般细胞稳态该仪器将使我们能够扩展。我们对与 NAA10 和 NAA15 相关疾病相关的分子生物学和病理生理学的研究和 NTA 途径，作为了解 NTA 在哺乳动物生物学中的作用的持续努力的一部分。研究将是揭示 NTA 在人类健康和疾病中的作用的关键一步，正如 NTA 一直在研究的那样与癌症进展和神经退行性疾病有关，包括帕金森氏症、阿尔茨海默氏症和亨廷顿舞蹈病。