Genetic and Molecular Analyses of Protein Biogenesis in the Neuroretina

神经视网膜蛋白质生物发生的遗传和分子分析

• 批准号: 8289556
• 负责人: PAULO A FERREIRA
• 金额: $ 37.44万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8289556
• 关键词: 26S proteasome; Achievement; Affect; Age; Aging; Ally; Binding Proteins; Biogenesis; Biological; Cells; Complex; Coupled; Cyclophilins; Disease; Ectopic Expression; Equilibrium; Ganglia; Goals; Homeostasis; Human; Impairment; Lead; Life; Light; Link; Molecular; Molecular Genetics; Mus; Mutate; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Opsin; Oxidative Stress; Pathogenesis; Peptidylprolyl Isomerase; Phenotype; Photoreceptors; Pigment Epithelium; Process; Proteins; Proteolysis; Regulation; Research; Retina; Retinal; Retinal Cone; Retinal Degeneration; Retinal Diseases; Role; Running; Stress; Structure of retinal pigment epithelium; System; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; Ubiquitin; Up-Regulation; Vision; Vision Disorders; age related; cell type; chaperone machinery; interdisciplinary approach; loss of function; loss of function mutation; mouse model; multicatalytic endopeptidase complex; neuroprotection; new therapeutic target; novel; prevent; protein degradation; protein folding; protein function; public health relevance; retinal neuron; retinal rods; sex

DESCRIPTION (provided by applicant): The deregulation of the ubiquitin-proteasome system (UPS) is a hallmark feature of many neurodegenerative disorders, including retinal degenerations, which involve misfolding and deregulation of protein turnover in photoreceptors. Our long-range objective is to understand the role of a regulatory complex of the UPS- chaperone machinery in controlling substrate degradation and folding and use this information to identify targets for novel therapeutic strategies, which presently lack efficacy toward selective biological and disease processes of the UPS. The goal of this proposal is to understand the molecular and intracellular mechanisms that sustain the balance between turnover and biogenesis of proteins, two tightly coupled processes that are vital for retinal-neuron function and survival and that are implicated in the pathogenesis of a wide variety of neuro-visual disorders of the human. Achievement of this goal will allow us and others to identify novel pharmacological targets and develop therapeutic strategies to manipulate the UPS-chaperone machinery, prevent the toxic deregulation of normal or expression of mutated proteins under various retinal disease states and aging manifestations causing the impairment of the UPS-chaperone machinery and neuro-visual function. The proposed research builds upon our prior research findings which demonstrated that substrates are presented to the proteasome upon synthesis and that the activity of the proteasome is regulated and compartmentalized in the cell. Ran-binding protein 2 (RanBP2) via two distinct domains, the cyclophilin-like (CLD) and cyclophilin (CY) domains, is a key controller of such processes. This proposal tests our overall hypothesis that RanBP2 and its CLD and CY domains control two tightly coupled processes, the UPS-dependent turnover and biogenesis of selective proteins that are critical to the function and survival of selective retinal neurons and retinal pigment epithelium (RPE) cells under normal and pathophysiological conditions. We will test this hypothesis by accomplishing the following specific aims, which focus on the mechanistic roles of RanBP2, and its CLD and CY domains, in the regulation of protein homeostasis and survival of selective retinal neurons under normal and disease stress conditions. Aim 1. Test the hypothesis that loss-of-function of RanBP2 has differential biological and pathological effects among retinal neurons, some of which may be essential to the viability or protein homeostasis of such neurons. Aim 2. Test the hypothesis that selective loss-of-function of the CLD activity of RanBP2 up-regulates selective functions of the UPS that are critical to homeostasis of selective retinal proteins, function and survival of retinal neurons and RPE cells. Aim 3. Test the hypothesis that the cis-trans prolyl isomerase (PPIase) activity in the CY domain of RanBP2 controls the biogenesis of selective proteins that are critical to the function or survival of retinal neurons and RPE cells. PUBLIC HEALTH RELEVANCE: The modulation of protein degradation and biogenesis in the retina is of vital importance to its function. Impairment of these processes is known to underlie the pathogenesis of several degenerative and aging disorders affecting retinal neurons. The proposal aims at elucidating the role of Ran-binding protein 2 in protein homeostasis and allied pathohysiological states in the retina.

描述（由申请人提供）：泛素蛋白蛋白酶体系统（UPS）的放松管制是许多神经退行性疾病的标志性特征，包括视网膜退化，涉及光感受器中蛋白质更换的错误折叠和失调。我们的远距离目标是了解UPS-伴侣机械的调节复合物在控制底物降解和折叠中的作用，并使用这些信息来识别新型治疗策略的目标，目前缺乏对UPS的选择性生物学和疾病过程的功效。该提案的目的是了解维持蛋白质营业额和生物发生之间平衡的分子和细胞内机制，这两个紧密耦合的过程对于视网膜 - 神经元功能和生存至关重要，并且与人类各种神经性疾病的发病机理有关。实现这一目标将使我们和其他人能够确定新颖的药理学靶标并制定治疗策略来操纵UPS-核心机械，防止在各种视网膜疾病状态下对正常的毒理或突变蛋白的表达以及衰老表现出现的突变蛋白的表达，从而导致UPS-Chaperone机械机械和神经机械和神经疗法的障碍。提出的研究基于我们先前的研究发现，该发现表明底物在合成后将底物呈现给蛋白酶体，并且蛋白酶体的活性受到细胞中的调节和分裂。 RAN结合蛋白2（RANBP2）通过两个不同的结构域分别是环粒蛋白样（CLD）和环磷脂（CY）结构域，是此类过程的关键控制器。该提案检验了我们的总体假设，即RANBP2及其CLD和CY结构域控制了两个紧密耦合的过程，在正常和病理生理学条件下，选择性蛋白的UPS依赖性周转和选择性蛋白的生物发生对选择性视网膜神经元和视网膜色素（RPE）细胞的功能和存活至关重要。我们将通过实现以下特定目标来检验该假设，该目标的重点是RANBP2的机械作用及其CLD和CY域在蛋白质稳态调节中以及在正常和疾病胁迫条件下选择性视网膜神经元的存活。 AIM 1。检验以下假设：RANBP2的功能丧失在视网膜神经元之间具有差异性生物学和病理作用，其中一些可能对此类神经元的生存力或蛋白质稳态至关重要。 AIM 2。测试RANBP2的CLD活性的选择性丧失的假设上调UPS的选择性功能对选择性视网膜蛋白，视网膜神经元和RPE细胞的功能和存活至关重要。 AIM 3。检验以下假设：RANBP2 Cy域中的顺式反应蛋白蛋白酶（PPIASE）活性控制了选择性蛋白的生物发生，这对于视网膜神经元和RPE细胞的功能或存活至关重要。 公共卫生相关性：视网膜中蛋白质降解和生物发生的调节对于其功能至关重要。已知这些过程的损害是影响视网膜神经元的几种退化性和衰老疾病的发病机理的基础。该提案旨在阐明ran结合蛋白2在视网膜中蛋白质稳态和盟军病原体状态中的作用。