ATase1 and ATase2, proteostasis, and neurological diseases

ATase1 和 ATase2、蛋白质稳态和神经系统疾病

基本信息

批准号：
10554962
负责人：
Luigi Puglielli
金额：
$ 30.03万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10554962
关键词：
Acetyl Coenzyme A Acetylation Acetyltransferase Animals Autophagocytosis Biochemical Biological Process Biology Brain Cellular biology Cessation of life Collaborations Communication Deacetylase Defect Developmental Delay Disorders Endoplasmic Reticulum Ensure Equilibrium Event Functional disorder Gene Duplication Genetic Glutamate-ammonia-ligase adenylyltransferase Glycoproteins Goals Golgi Apparatus Heterozygote Human Intellectual functioning disability Laboratories Lysine Maintenance Mass Spectrum Analysis Mediating Membrane Transport Proteins Metabolic Modeling Molecular Molecular Biology Mus Mutation Neurons Neuropathy Organelles Output Pathway interactions Peripheral Phenotype Progeria Proteins Quality Control Research Series Techniques Testing Therapeutic autism spectrum disorder design disease phenotype experimental study human disease in vivo loss of function mouse model nervous system disorder novel pharmacologic premature protein aggregation proteostasis response trafficking translational approach

项目摘要

We discovered that Nε-lysine acetylation occurs in the lumen of the endoplasmic reticulum (ER) in 2007. From that initial finding, we went on to discover the entire ER acetylation machinery (one membrane transporter, AT-1/SLC33A1, and two acetyltranferases, ATase1 and ATase2) and uncover a novel piece of ER biology. Specifically, we discovered that the ER acetylation machinery regulates proteostasis within the secretory pathway as well as metabolic crosstalk between different intracellular organelles and compartments. Human-based studies discovered that dysfunctional ER acetylation, as caused by loss-of-function homozygous and heterozygous mutations or gene duplication events, is associated with different human diseases, from developmental delay of the brain and premature death to peripheral forms of neuropathy, autism spectrum disorder, intellectual disability and segmental progeria. Mouse models that mimic these genetic events recapitulate associated human diseases. Importantly, ATase-targeting compounds that restore the proteostatic functions of the ER rescue the disease phenotypes of the animals. In conclusion, we have identified a novel molecular machinery that is key to the maintenance of proteostasis within the secretory pathway, and that can be targeted to (i) understand the pathophysiology of several related neurological diseases and (ii) develop appropriate translational approaches to resolve proteostatic defects. The GENERAL HYPOTHESIS of this research is that ATase1 and ATase2 act downstream of an intracellular communication network that regulates the proteostatic functions of the ER and secretory pathway. Our main goal is to dissect the molecular mechanism(s) underlying the acetyl-CoA:lysine acetyltransferase activity of the ATases and understand how ER-based acetylation regulates the efficiency of the secretory pathway. Aim 1 will test the hypothesis that the ATases have divergent functions and differentially regulate proteostasis and metabolic crosstalk. Aim 2 will test the hypothesis that unique structural features allow the ATases to respond to acetyl-CoA influx, Ca++ levels, and perturbations in ER proteostasis. Aim 3 will test the hypothesis that the acetyl group added in the ER lumen by the ATases must be removed in the lumen of the Golgi apparatus by Amfion/GDAC to ensure correct trafficking of nascent glycoproteins and the quality of the secretome. In conclusion, this proposal is based on novel findings from our laboratory and offers a series of highly mechanistic studies that have the potential to define new avenues of research (and treatment) for different neurological diseases. The proposal will use unique mouse models as well as highly integrated novel experimental approaches. We believe that upon completion of these studies, we will have defined an entirely new avenue of research for different neurological diseases.

2007 年，我们发现 Nε-赖氨酸乙酰化发生在内质网 (ER) 的腔中。从最初的发现开始，我们继续发现了整个 ER 乙酰化机制（一个膜转运蛋白 AT-1/SLC33A1 和两个乙酰转移酶））、ATase1 和 ATase2）并揭示了 ER 生物学的一个新部分具体来说，我们发现了 ER 乙酰化机制。调节分泌途径内的蛋白质稳态以及不同细胞内细胞器和区室之间的代谢串扰基于人类的研究发现，由功能丧失的纯合子和杂合子突变或基因复制事件引起的功能失调的 ER 乙酰化与不同的人类有关。疾病，从大脑发育迟缓和过早死亡到周围神经病、自闭症谱系障碍、智力障碍和节段性早衰症，模拟这些遗传事件的小鼠模型再现了相关的人类。重要的是，恢复内质网蛋白抑制功能的 ATase 靶向化合物可以挽救动物的疾病表型。总之，我们已经确定了一种新的分子机制，它是维持分泌途径中蛋白质稳态的关键，并且可以针对（i）了解几种相关神经系统疾病的病理生理学和（ii）开发适当的转化方法来解决本研究的一般假设是 ATase1 和 ATase2 作用于调节 ER 和分泌途径的蛋白抑制功能的细胞内通讯网络的下游。剖析 ATase 的乙酰辅酶 A：赖氨酸乙酰转移酶活性的分子机制，并了解基于 ER 的乙酰化如何调节分泌途径的效率。目标 1 将检验 ATase 具有不同功能并差异调节蛋白质稳态和代谢串扰的假设，目标 2 将检验独特的结构特征允许 ATase 对乙酰辅酶 A 流入、Ca++ 水平和 ER 蛋白质稳态扰动做出反应的假设。目标 3 将检验以下假设：由 ATase 在 ER 腔中添加的乙酰基必须在 ER 腔中去除。 Amfion/GDAC 的高尔基体确保新生糖蛋白的正确运输和分泌蛋白组的质量。总之，该提案基于我们实验室的新发现，并提供了一系列高度机械化的研究，有可能为不同的神经系统疾病定义新的研究（和治疗）途径。该提案将使用独特的小鼠模型以及。我们相信，完成这些研究后，我们将为不同的神经系统疾病定义一条全新的研究途径。