Dendritic spine morphology and the Ubiquitin pathway

树突棘形态和泛素通路

• 批准号: 7388865
• 负责人: Xiaolu Ang Cambronne
• 金额: $ 2.53万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7388865
• 关键词: Actins; Acute; Affect; Binding; Biological Assay; Cells; Classification; Complex; Computer software; Cytoskeleton; DNA Sequence Rearrangement; Data; Dendritic Spines; Disease; Dominant-Negative Mutation; Down Syndrome; Down-Regulation; Epilepsy; Excitatory Synapse; F-Actin; F-Box Proteins; Fragile X Syndrome; Green Fluorescent Proteins; Head; Hippocampus (Brain); Image; In Vitro; Individual; Investigation; Ligase; Mediating; Molecular; Morphology; N-Methyl-D-Aspartate Receptors; Neurons; Numbers; Pathway interactions; Phosphorylation; Picrotoxin; Proteasome Inhibitor; Protein Overexpression; Protein-Serine-Threonine Kinases; RNA Interference; Reaction; Recruitment Activity; Regulation; Role; SKP Cullin F-Box Protein Ligases; Signal Transduction; Specificity; Structure; Synapses; Synaptic plasticity; TP53 gene; Ubiquitin; Ubiquitination; Vertebral column; base; density; genetic regulatory protein; human PLK1 protein; in vivo; knock-down; nervous system disorder; p27 Cell Cycle Protein; p27 Enzyme Inhibitor; postsynaptic; presynaptic density protein 95; reconstitution; research study; serum-inducible kinase; size; two-photon; ubiquitin-protein ligase

Dendritic spines in excitatory synapses can undergo dynamic actin-based morphological changes associated with synaptic plasticity. Recent examination into the molecular basis of this plasticity uncovered a role for SPAR, a RapGAP that targets to the PSD of dendritic spines. In turn, SPAR stability is regulated by serine/threonine kinase SNK. SNK phosphorylates SPAR, which promotes SPAR degradation through a proteasomal pathway. Nevertheless, the identity of the E3 Ub-ligases that target SPAR and how they are regulated is still unknown. Our investigation into the E3 Ub-ligases that target SPAR to regulate synaptic plasticity includes the following three major efforts: Aim 1 investigates roles for SCF Ub-ligases in SPAR degradation and the activation signals they recognize in the substrate; Aim 2 details a comprehensive approach to identify all E3 Ub-ligases that target SPAR; and Aim 3 examines how depletion of identified candidate E3 Ub-ligases affects dendritic spine morphology in primary hippocampal neurons. In addition to providing concrete evidence for the involvement of the Ub-pathway in the regulation of dendritic spine morphology, identification of the E3 Ub-ligases that mediate SPAR degradation is intrinsic to understanding the mechanism that may underlie a number of disorders associated with abnormal spine morphology and density, including general mental retardation, fragile-X syndrome, Down's syndrome, and epilepsy.

兴奋性突触中的树突棘可以经历基于肌动蛋白的动态形态变化 具有突触可塑性。最近对这种可塑性的分子基础的研究发现了 SPAR，一种针对树突棘 PSD 的 RapGAP。反过来，SPAR 稳定性受以下因素调节： 丝氨酸/苏氨酸激酶 SNK。 SNK 磷酸化 SPAR，从而通过 蛋白酶体途径。然而，针对 SPAR 的 E3 Ub 连接酶的身份及其作用方式 是否受到监管仍是未知数。我们对靶向 SPAR 来调节突触的 E3 Ub 连接酶的研究 可塑性包括以下三项主要工作： 目标 1 研究 SCF Ub 连接酶在 SPAR 中的作用 降解和它们在基质中识别的激活信号；目标 2 详细介绍了全面的 识别所有针对 SPAR 的 E3 Ub 连接酶的方法；目标 3 检查已识别的物质的消耗如何 候选 E3 Ub 连接酶影响原代海马神经元的树突棘形态。此外 为Ub通路参与树突棘调控提供具体证据 形态学方面，介导 SPAR 降解的 E3 Ub 连接酶的鉴定对于理解至关重要 可能是许多与脊柱形态异常相关的疾病的机制 密度，包括一般精神发育迟滞、脆性 X 综合征、唐氏综合症和癫痫。