Investigating the functional consequences of SynGAP1 SUMOylation at mammalian synapses

研究哺乳动物突触中 SynGAP1 SUMO 化的功能后果

• 批准号: EP/Y024559/1
• 负责人: Marie Pronot
• 金额: $ 23.84万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FY024559%2F1
• 关键词: Investigating; functional; consequences; SynGAP1; SUMOylation

ntellectual disability (ID) affects millions of individuals worldwide and represents a major health and economic burden. ID results from mutations altering the function of different brain proteins. One of these proteins is SynGAP1. In humans, truncated or missense variants in the SYNGAP1 gene are associated with ID, epilepsy and autism spectrum disorders. SynGAP1 is critical for synapse formation and morphology and is a negative regulator of excitatory neurotransmission. Furthermore, the post-translational modification process of SUMOylation is emerging as a key regulator of synapse development and function. I have key pilot data showing that SynGAP1 is a SUMO target at synapses. This raises the hypothesis that SUMOylation regulates SynGAP1 activity and function, to play a pivotal role in synapse formation, morphology and function. To unveil the physiological consequences of SynGAP1 SUMOylation on its molecular function, I will determine which signaling cascades mediate its SUMOylation, and how this impacts its enzyme activity, molecular interactions and subcellular localisation. To determine effects on synapse morphology and neurotransmission (which are controlled by SynGAP1 and usually altered in ID), I will exploit live-cell imaging and electrophysiological approaches with sophisticated biochemical assays in Syngap1-/- neurons that express a form of SynGAP that cannot bind to SUMO (SynGAP1-K149R). Therefore, the proposed project will exploit state of the art technologies to directly address the molecular mechanisms regulating SynGAP1 function in neurons and the synaptic events it controls. Furthermore, it will reveal essential insights regarding the role of SUMO in the development of ID. This is an emerging issue of critical importance, since several proteins essential for correct brain development and function are SUMO targets. Therefore, this project will also facilitate future strategies to restore disrupted synaptic function by targeting SUMO.ntellectual disability (ID) affects millions of individuals worldwide and represents a major health and economic burden. ID results from mutations altering the function of different brain proteins. One of these proteins is SynGAP1. In humans, truncated or missense variants in the SYNGAP1 gene are associated with ID, epilepsy and autism spectrum disorders. SynGAP1 is critical for synapse formation and morphology and is a negative regulator of excitatory neurotransmission. Furthermore, the post-translational modification process of SUMOylation is emerging as a key regulator of synapse development and function. I have key pilot data showing that SynGAP1 is a SUMO target at synapses. This raises the hypothesis that SUMOylation regulates SynGAP1 activity and function, to play a pivotal role in synapse formation, morphology and function. To unveil the physiological consequences of SynGAP1 SUMOylation on its molecular function, I will determine which signaling cascades mediate its SUMOylation, and how this impacts its enzyme activity, molecular interactions and subcellular localisation. To determine effects on synapse morphology and neurotransmission (which are controlled by SynGAP1 and usually altered in ID), I will exploit live-cell imaging and electrophysiological approaches with sophisticated biochemical assays in Syngap1-/- neurons that express a form of SynGAP that cannot bind to SUMO (SynGAP1-K149R). Therefore, the proposed project will exploit state of the art technologies to directly address the molecular mechanisms regulating SynGAP1 function in neurons and the synaptic events it controls. Furthermore, it will reveal essential insights regarding the role of SUMO in the development of ID. This is an emerging issue of critical importance, since several proteins essential for correct brain development and function are SUMO targets. Therefore, this project will also facilitate future strategies to restore disrupted synaptic synaptic function by targeting SUMO.

ntelectual disability（id）影响了全球数百万个人，代表了重大的健康和经济负担。 ID是由于突变改变了不同脑蛋白的功能。这些蛋白质之一是syngap1。在人类中，Syngap1基因中的截短或错义变体与ID，癫痫和自闭症谱系障碍有关。 Syngap1对于突触形成和形态至关重要，并且是兴奋性神经传递的负调节剂。此外，Sumoylation的翻译后修饰过程正在成为突触发展和功能的关键调节剂。我有关键的试点数据，显示Syngap1是突触下的Sumo目标。这提出了一个假设，即Sumoylation调节Syngap1活性和功能，以在突触形成，形态和功能中起关键作用。为了揭示Syngap1 sumoylation对其分子功能的生理后果，我将确定哪些信号级联反应介导其Sumoylation，以及这如何影响其酶活性，分子相互作用和亚细胞定位。为了确定对突触形态和神经传递的影响（由Syngap1控制并通常在ID中更改），我将使用Syngap1 - / - / - 神经元中的生物化学测定来利用活细胞成像和电生理方法，这些神经元在表达合成形式的Sumo（Syngap1-kap1-k1-1-1-1-1-149r）中表达合成形式的形式。因此，拟议的项目将利用艺术技术的状态直接解决调节神经元中Syngap1功能的分子机制及其控制的突触事件。此外，它将揭示有关Sumo在ID发展中的作用的基本见解。这是一个至关重要的新兴问题，因为几种对于正确的大脑发育和功能所必需的蛋白质是相扑靶标。因此，该项目还将促进未来的策略来通过瞄准Sumo来恢复突触功能的破坏。 ID是由于突变改变了不同脑蛋白的功能。这些蛋白质之一是syngap1。在人类中，Syngap1基因中的截短或错义变体与ID，癫痫和自闭症谱系障碍有关。 Syngap1对于突触形成和形态至关重要，并且是兴奋性神经传递的负调节剂。此外，Sumoylation的翻译后修饰过程正在成为突触发展和功能的关键调节剂。我有关键的试点数据，显示Syngap1是突触下的Sumo目标。这提出了一个假设，即Sumoylation调节Syngap1活性和功能，以在突触形成，形态和功能中起关键作用。为了揭示Syngap1 sumoylation对其分子功能的生理后果，我将确定哪些信号级联反应介导其Sumoylation，以及这如何影响其酶活性，分子相互作用和亚细胞定位。为了确定对突触形态和神经传递的影响（由Syngap1控制并通常在ID中更改），我将使用Syngap1 - / - / - 神经元中的生物化学测定来利用活细胞成像和电生理方法，这些神经元在表达合成形式的Sumo（Syngap1-kap1-k1-1-1-1-1-149r）中表达合成形式的形式。因此，拟议的项目将利用艺术技术的状态直接解决调节神经元中Syngap1功能的分子机制及其控制的突触事件。此外，它将揭示有关Sumo在ID发展中的作用的基本见解。这是一个至关重要的新兴问题，因为几种对于正确的大脑发育和功能所必需的蛋白质是相扑靶标。因此，该项目还将促进未来的策略，以瞄准SUPO，以恢复突触突触功能。