Structural and Molecular Mechanisms for Dysregulation of Protein Kinase C Gamma in Cerebellar Ataxia

小脑性共济失调中蛋白激酶 C γ 失调的结构和分子机制

基本信息

批准号：
10394960
负责人：
ALEXANDRA C. NEWTON
金额：
$ 65.3万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10394960
关键词：
Address Affect Ataxia Automobile Driving Biochemical Brain Cerebellar Ataxia Cerebellar Diseases Cerebellum Cryoelectron Microscopy Cyclic AMP-Dependent Protein Kinases Diglycerides Disease Enzymes Fluorescence Resonance Energy Transfer Future Germ-Line Mutation Goals Homeostasis Isoenzymes Knowledge Length Malignant Neoplasms Molecular Molecular Conformation Motor Mutation Neurodegenerative Disorders Neurons Output Pathology Pathway interactions Patients Phosphotransferases Protein Region Proteins Public Health Purkinje Cells Quality Control Regulation Research Resolution Shunt Device Signal Pathway Signal Transduction Signaling Protein Spinocerebellar Ataxias Spinocerebellar Degenerations Structure Therapeutic Vision disease phenotype live cell imaging mouse model mutant novel phosphoproteomics protein function protein kinase C gamma sensor spatiotemporal structural biology transcriptome transcriptome sequencing

项目摘要

Summary/Abstract The overall vision of our proposed research is to understand the structural, molecular, and cellular mechanisms by which germline mutations in protein kinase C gamma (PKCg) drive the neuro- degenerative disease Spinocerebellar Ataxia 14 (SCA14). PKCg is a Ca2+/diacylglycerol-regulated kinase expressed only in neurons, including Purkinje cells whose degeneration is a hallmark of the almost 50 subtypes of SCA. We have assembled a team with extensive and complementary expertise in structural biology of kinases and in PKC mechanisms to understand how these mutations alter the structure and function of PKCg to contribute to the disease phenotype. The hypothesis driving this proposal is that mutations are concentrated at specific regions of PKCg that break autoinhibitory contacts to enhance its activity by a novel mechanism that evades normal quality control degradation. This evasion of degradation may be a unique feature of the Ataxia mutations as cancer-associated mutations that break autoinhibitory contacts destabilize PKC and shunt it to degradation. Such evasion of normal quality control allows aberrantly active PKCg to enhance its signaling output, which in Purkinje cells in the cerebellum leads to degeneration. Furthermore, we hypothesize that enhanced signaling by PKCg may underlie the pathology of SCA, in general, as a large fraction of SCAs are caused by mutations in proteins that control Ca2+ homeostasis or signaling. We aim to combine computational, structural, biochemical, live-cell imaging, and phosphoproteomics approaches to understand the molecular details of how disease-associated mutations in PKCg impact function, with the long-term future goal of using this knowledge to treat this devastating disease.

摘要/摘要我们提出的研究的总体视野是了解结构，分子和细胞蛋白激酶C伽马（PKCG）中种系突变驱动神经的机制退化性疾病脊椎园共济失调14（SCA14）。 PKCG是Ca2+/二酰基甘油调节的激酶仅在神经元中表达，包括浦肯野细胞，其变性是近50个的标志 SCA的亚型。我们已经组建了一个在结构上具有广泛和互补专业知识的团队激酶和PKC机制的生物学，以了解这些突变如何改变结构和 PKCG有助于疾病表型的功能。推动该提议的假设是突变集中在破裂自身抑制性接触的PKCG的特定区域，以增强其通过逃避正常质量控制降解的新型机制的活性。这种逃避降解可能是共济失调突变的独特特征，因为癌症相关突变破裂自动抑制性接触不稳定PKC并将其分流至降解。这种逃避正常质量控制允许异常活跃的PKCG增强其信号输出，该输出在Purkinje细胞中小脑会导致变性。此外，我们假设PKCG的增强信号传导可能通常，SCA的病理基础，因为大量SCA是由蛋白质突变引起的控制CA2+稳态或信号传导。我们旨在结合计算，结构，生化，活细胞成像和磷酸蛋白质组学方法来了解如何 PKCG撞击功能中与疾病相关的突变，其长期未来的目标是使用知识治疗这种毁灭性疾病。