Elucidating the Complexin C-terminal Domain Mechanism in Neurotransmission Regulation

项目总结 复杂蛋白C末端结构域在神经递质调节中的作用机制 神经传递的失调是阿尔茨海默氏症等疾病的神经病理生理学基础， 帕基森氏病、肌萎缩侧索硬化症和自闭症。突触小泡(SV)融合是一种 神经传递过程和SNARE(可溶性NSF附着蛋白受体)蛋白是必不可少的 与SV融合有关，但SNARS的调节蛋白是了解其机制的关键。尽管 圈套发现以来奇妙的研究进展，驱动SV融合的机制细节 仍然难以捉摸。复合素(CPX)是SNARs的调节器，通过曲率传感定位于SVS 基序位于其C-末端结构域(CTD)。引人注目的是，cpx在动物界是高度保守的--甚至 像Trichoplax这样缺乏真正突触的简单多细胞生物有CPX。线虫借给自己 作为研究环磷酰胺生物学的理想模式生物，环磷酰胺零突变具有致命性。 此外，线虫很容易想象，并提供了一种强大的遗传工具，其起源是一种 遗传学模型生物体。 据报道，环磷酰胺的变异体可引起婴儿肌阵挛癫痫和智力障碍。 残疾(Redler，S.等人)2017欧元J Hum Genet)。具体地说，CTD发生了突变，效率低下 复合蛋白的定位被认为是导致疾病的原因。我们的研究计划调查CTD是否 在功能上对复杂蛋白至关重要，以及复杂蛋白CTD在将其本地化到 突触小泡。在目标1中，我将探讨CPX-1 CTD在特定水泡池招募中的作用 将突触与靶CPX-1的N端的一半结合，用外源绳索绕过CTD膜结合。在……里面 目标2，我将描述有效的SV定位所需的CTD曲率传感器的特性和 探索CPX-1和SV蛋白RAB-3之间潜在的生化相互作用。我还将探索 在cpx-1的正向遗传抑制筛查中发现的几个基因，以扩大我对潜在cpx-1的搜索 1结合伙伴。这些目标将结合遗传、分子、成像和生化方法。 剖析和刻画CPX的一个关键区域，回答一个长期存在的问题 CPX的突触机制。 在这份奖学金下，我将有机会与三所学院校区的主要研究人员合作 (威尔·康奈尔、洛克菲勒和斯隆·凯特林纪念馆)进行神经科学和生物物理学研究 在一个协作和支持的环境中。为了扩展我的技术技能和知识，我将参加 关于对我的研究重要的主题的研讨会、研讨会和会议。研究成果将被分享 通过会议和出版物与科学界和公众进行交流。

PROJECT SUMMARY Elucidating the Complexin C-terminal domain mechanism in neurotransmission regulation Dysregulation of neurotransmission underlies the neuropathophysiology of conditions like Alzheimer’s, Parkisons Disease, amyotrophic lateral sclerosis, and autism. Synaptic vesicle (SV) fusion is a central process in neurotransmission and SNARE (Soluble NSF Attachment Protein Receptor) proteins are essential to SV fusion, but the regulatory proteins of SNAREs are key to understanding the mechanisms. Despite fantastic research progress since the discovery of SNAREs, the mechanistic details driving SV fusion remain elusive. Complexin (Cpx) is a regulator of SNAREs and localizes to SVs via a curvature-sensing motif in its C-terminal domain (CTD). Strikingly, Cpx is highly conserved in the animal kingdom – even simple multicellular organisms lacking bona fide synapse like Trichoplax have Cpx. C. elegans lends itself as an ideal model organism to study Cpx biology as the mammalian Cpx null mutation is lethal. Furthermore, C.elegans is easy to image and provides a powerful genetic tool with its origins as a genetics model organism. Variants of Cpx have been reported to be pathogenic for infantile myoclonic epilepsy and intellectual disability (Redler, S. et al. Eur J Hum Genet 2017). Specifically, the CTD was mutated and the inefficient localization of complexin is thought to result in disease. Our research plan investigates whether the CTD is functionally essential to complexin and which mechanisms the complexin CTD employs in localizing to synaptic vesicles. In Aim 1, I will explore the role of the CPX-1 CTD in recruitment to specific vesicle pools at the synapse and target the N-terminal half of CPX-1 with foreign tethers to bypass CTD membrane binding. In Aim 2, I will characterize the features of the CTD curvature sensor required for efficient SV targeting and explore potential biochemical interactions between CPX-1 and the SV protein RAB-3. I will also explore several genes identified in a forward genetic suppressor screen of cpx-1 to extend my search of potential CPX- 1 binding partners. These aims will combine genetic, molecular, imaging, and biochemical approaches to dissect and characterize a critical region of Cpx, providing answers to a long-standing question on the synaptic mechanisms of Cpx. Under this fellowship, I will have the opportunity to work with leading researchers at the Tri-Institutional campus (Weill Cornell, Rockefeller, and Memorial Sloan Kettering) conducting neuroscience and biophysical research in a collaborative and supportive environment. To expand my technical skills and knowledge, I will attend workshops, seminars, and conferences on topics important for my research. Research findings will be shared with the scientific community and public via conferences and publications.