REGULATION OF INTRACELLULAR TRANSDUCIN HOMEOSTASIS BY PHOSDUCIN IN VERTEBRATE RO

磷酸酯蛋白对脊椎动物 RO 细胞内转导蛋白稳态的调节

• 批准号: 7719933
• 负责人: MAXIM SOKOLOV
• 金额: $ 16.84万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7719933
• 关键词: Address; Afferent Neurons; Blindness; Computer Retrieval of Information on Scientific Projects Database; Coupled; Detection; Dissection; Funding; GTP-Binding Proteins; Goals; Grant; Half-Life; Heterotrimeric GTP-Binding Proteins; Homeostasis; Immunoprecipitation; In Vitro; Institution; Knock-out; Knockout Mice; Laboratories; Life; Mediating; Messenger RNA; Molecular; Phosphorylation; Photoreceptors; Property; Proteins; Regulation; Reporting; Research; Research Design; Research Personnel; Resources; Retina; Retinal; Retinal Degeneration; Retinal Photoreceptors; Role; Signal Transduction; Source; Transducin; Transgenic Organisms; United States National Institutes of Health; Vertebrate Photoreceptors; Wild Type Mouse; base; beta Subunit Transducin; cell growth regulation; in vivo; mutant; phosducin; photoreceptor degeneration; retinal rods; visual information; visual stimulus; Address; Afferent Neurons; Blindness; Computer Retrieval of Information on Scientific Projects Database; Coupled; Detection; Dissection; Funding; GTP-Binding Proteins; Goals; Grant; Half-Life; Heterotrimeric GTP-Binding Proteins; Homeostasis; Immunoprecipitation; In Vitro; Institution; Knock-out; Knockout Mice; Laboratories; Life; Mediating; Messenger RNA; Molecular; Phosphorylation; Photoreceptors; Property; Proteins; Regulation; Reporting; Research; Research Design; Research Personnel; Resources; Retina; Retinal; Retinal Degeneration; Retinal Photoreceptors; Role; Signal Transduction; Source; Transducin; Transgenic Organisms; United States National Institutes of Health; Vertebrate Photoreceptors; Wild Type Mouse; base; beta Subunit Transducin; cell growth regulation; in vivo; mutant; phosducin; photoreceptor degeneration; retinal rods; visual information; visual stimulus

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Retinal photoreceptors, rods and cones, are highly specialized sensory neurons that utilize G protein-mediated signaling for acquisition of visual information. Rods maintain high, invariable amounts of heterotrimeric G protein transducin required for the detection of visual stimuli, throughout their lives. Importantly, drastic change of transducin intracellular amounts often puts photoreceptors on the path to elimination, which causes retina degeneration and loss of vision. The long-term goal of our laboratory is to understand the role of G protein-mediated signaling in retinal photoreceptor function. The objective of this application is to elucidate molecular mechanisms underlying transducin homeostasis in rods. The central hypothesis of this application is that transducin accumulation in rods is regulated by phosducin, a regulator of G protein mediated signaling, which undergoes a cycle of light-dependent phosphorylation. This hypothesis is based on the reported in vitro properties of phosducin and our observation that rods of the phosducin knockout mouse accumulate significantly reduced amount of transducin. In Aims 1 and 2 we will elucidate the role of phosducin in transducin transcriptional and proteolytic control. For that we will compare the transducin beta mRNA levels, as well as values of transducin beta protein half-life in the retinas of the phosducin knockout and wild type mice. Aim 3 will be dedicated to studies of light-dependent phosducin phosphorylation, where we will determine intracellular localization of the phosducin phospho-states within the rod, address the in vivo function of this regulation using transgenic expression of phosducin phosphorylation mutants, and identify protein partners of phosducin in various rod subcellular compartments using retinal micro-dissection and immunoprecipitation coupled to mass spectrometric protein identification. The proposed studies are designed to elucidate the role of phosducin in regulation of transducin homeostasis in photoreceptors, which is important for our understanding of the molecular mechanisms of photoreceptor degeneration and for developing strategies to counteract retinal degeneration. We expect that our results will also have a significant impact on our understanding of the principles of cellular regulation of G protein-mediated signaling.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 视网膜感受器，棒和锥体是高度专业的感觉神经元，利用G蛋白介导的信号传导来获取视觉信息。杆在其一生中保持高量的异三聚体G蛋白透明素。重要的是，细胞内量的急剧变化通常会使光感受器处于消除道路上，这会导致视网膜变性和视力丧失。 我们实验室的长期目标是了解G蛋白介导的信号传导在视网膜感受器功能中的作用。该应用的目的是阐明杆中跨丁质稳态的基础机制。该应用的中心假设是，杆中的透明蛋白积累受到G蛋白介导的信号传导的调节剂的调节，后者经历了光依赖性磷酸化的循环。该假设是基于报道的硫糖素的体外特性，我们的观察结果是，哲学敲除小鼠的杆会显着降低转丁蛋白的量。在AIMS 1和2中，我们将阐明磷酸素在转杜蛋白转录和蛋白水解对照中的作用。为此，我们将比较phosducin敲除和野生型小鼠的视网膜中透明蛋白βmRNA水平以及透明蛋白β蛋白半衰期的值。目标3将致力于研究光依赖性磷酸磷酸化的研究，在此研究中，我们将确定在杆中的磷酸磷酸化 - 磷酸化态的细胞内定位，以这种调控的体内功能，利用磷酸化磷酸化突变体中的磷酸化磷酸化蛋白质的磷酸化蛋白质的磷酸化蛋白质的磷酸化蛋白质的磷酸化蛋白质的磷酸化蛋白质的体内功能结合质谱蛋白鉴定。 拟议的研究旨在阐明磷素在光感受器中调节透明蛋白稳态中的作用，这对于我们对感光体变性的分子机制以及制定抵抗视网膜变性的策略很重要。我们期望我们的结果还将对我们对G蛋白介导的信号传导细胞调节原理的理解产生重大影响。