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愿景是了解结构、分子和细胞蛋白激酶 C γ (PKCg) 种系突变驱动神经细胞的机制退行性疾病脊髓小脑共济失调 14 (SCA14)。 PKCg 是一种 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，其退化是近 50 SCA 的亚型。我们组建了一支在结构领域拥有广泛且互补的专业知识的团队激酶生物学和 PKC 机制，以了解这些突变如何改变结构和 PKCg 的功能有助于疾病表型。推动该提议的假设是突变集中在 PKCg 的特定区域，破坏自抑制接触以增强其通过避免正常质量控制退化的新颖机制来进行活性。这种逃避降解可能是共济失调突变的一个独特特征，因为癌症相关突变会破坏自抑制接触会破坏 PKC 的稳定性并使其降解。这种逃避正常品质的行为控制允许异常活跃的 PKCg 增强其信号输出，这在浦肯野细胞中小脑导致退化。此外，我们假设 PKCg 增强的信号传导可能一般而言，SCA 是 SCA 病理学的基础，因为大部分 SCA 是由蛋白质突变引起的控制 Ca2+ 稳态或信号传导。我们的目标是将计算、结构、生物化学、活细胞成像和磷酸蛋白质组学方法来了解分子细节与疾病相关的 PKCg 突变会影响功能，未来的长期目标是利用该突变治疗这种毁灭性疾病的知识。