Cellular mechanisms of nucleocytoplasmic export through Nuclear Envelope Budding

通过核膜出芽的核细胞质输出的细胞机制

• 批准号: 10655419
• 负责人: SUSAN M PARKHURST
• 金额: $ 38.66万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-03 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655419
• 关键词: Actins; Aging; Biochemical; Biological; Bypass; Caring; Cell Nucleus; Cell Proliferation; Cell physiology; Cells; Clinical; Complex; Cytomegalovirus Infections; Cytoplasm; Development; Developmental Process; Disease; Drosophila genus; Elements; Encapsulated; Engineering; Event; Family; Foundations; Genes; Genetic; Goals; Growth; Herpesviridae; Herpesviridae Infections; Image; Immune; Immune system; Intervention; Kinesin; Learning; Malignant Neoplasms; Mammalian Cell; Mechanics; Mediating; Membrane; Microscopy; Molecular; Movement; Nuclear; Nuclear Envelope; Nuclear Export; Nuclear Inner Membrane; Nuclear Lamina; Nuclear Outer Membrane; Nuclear Pore; Nuclear Pore Complex; Nucleic Acids; Oogenesis; Organism; Pathway interactions; Physiological; Predisposition; Process; Proteins; Proteomics; RNA; Reagent; Regulation; Ribonucleoproteins; Role; Symptoms; Syndrome; System; Techniques; Transplant Recipients; Transport Process; Viral; Wiskott-Aldrich Syndrome; Work; clinically relevant; design; experimental study; information gathering; insight; interest; macromolecule; nervous system disorder; new therapeutic target; novel; nucleocytoplasmic transport; optogenetics; spatiotemporal; superresolution imaging; synaptogenesis; therapeutic development; trafficking; tumor; tumor progression; ultra high resolution

PROJECT SUMMARY/ABSTRACT Transport of nucleic acids and proteins from the nucleus to the cytoplasm is essential for nearly all cellular processes, and when mis-regulated, is associated with diseases, tumor formation/growth, and cancer progression. Canonically, this indispensable process has been thought to occur exclusively via Nuclear Pore Complexes, which span the nuclear envelope’s double membranes and provide a critical regulatory step in what exits (and enters) the nucleus. Recently, Nuclear Envelope (NE-) budding was shown to provide an alternative pathway for nuclear exit, particularly for large ribonucleoprotein (RNP) complexes that would otherwise need to unfold/remodel to fit through the pores. In this pathway, large macromolecule complexes are encapsulated by the inner nuclear membrane, cross the perinuclear space, fuse with the outer nuclear membrane, and are released into the cytoplasm, a mechanism strikingly similar to herpesvirus nuclear egress. Thus, NE-budding elegantly allows for large RNP complexes to exit the nucleus together and be delivered as a package for specific cellular functions. Despite its clear biological importance and clinical relevance, very little is yet known about the regulatory or structural machineries that allow NE-budding to occur in any system. Recently, we found that the Wiskott Aldrich Syndrome family actin nucleation protein, WASH, its four subunit regulatory complex (SHRC), and Arp2/3 are necessary for NE-budding. Using WASH/SHRC as a new entry point, in tandem with strategies to discover novel genes/proteins involved in this process, our long-term goal is to understand the molecular and cellular mechanics that govern NE-budding. The specific aims of this proposal are to determine the mechanism(s) of WASH/SHRC function in NE-budding, and to identify/analyze the infrastructural components/machineries governing the dynamic NE-budding process using a combination of genetic, biochemical, cell biological, time-lapse live imaging, and super- resolution/EM microscopy approaches. Drosophila provides an excellent, genetically amenable, organism for studying this conserved process due to its amenability for imaging and the wealth of cutting edge cell/molecular techniques and reagents. The information gathered in these studies will help to elucidate the mechanisms governing this exciting new nuclear export pathway in normal development or when mis-regulated in disease conditions, and may inform the study of herpesvirus nuclear egress as well.

项目摘要/摘要 核酸和蛋白质从细胞核到细胞质的运输对几乎所有细胞都是必不可少的。 过程，以及当调控不当时，与疾病、肿瘤形成/生长和癌症有关 进步。按照惯例，这一不可或缺的过程被认为完全是通过核能进行的 孔道复合体，跨越核膜的双层膜，提供关键的调节 步入退出(和进入)核团的部分。最近，核膜(NE-)萌发被证明是 为核退出提供替代途径，特别是对于大的核糖核蛋白(RNP)复合体 否则需要展开/重塑才能穿过毛孔。在这条道路上，大型 大分子复合体被内核膜包裹，穿过核周 空间，与外核膜融合，并被释放到细胞质中，这是一种机制 与疱疹病毒核出口惊人地相似。因此，东北萌芽优雅地允许较大的RNP 复合体一起退出细胞核，并作为特定细胞功能的包裹递送。 尽管它具有明显的生物学重要性和临床相关性，但人们对其调控机制知之甚少。 或允许在任何系统中发生东北萌芽的结构机械。最近，我们发现， Wiskott Aldrich综合征家族肌动蛋白核蛋白，WASH，其四个亚单位调节复合体 (SHRC)和Arp2/3是东北出芽所必需的。同时使用WASH/SHRC作为新的切入点 通过发现参与这一过程的新基因/蛋白质的策略，我们的长期目标是 了解控制东北萌发的分子和细胞机制。这样做的具体目的是 建议确定WASH/SHRC在NE萌发中的作用机制(S)，并 确定/分析管理动态NE萌芽过程的基础设施组件/机制 使用遗传学、生化、细胞生物学、延时实时成像和超 分辨率/EM显微镜接近。果蝇提供了一种极好的，遗传上顺从的， 用于研究这一保守过程的有机体，因为它适合成像和丰富的切割 边缘细胞/分子技术和试剂。在这些研究中收集的信息将有助于 阐明这一令人兴奋的新的核出口途径在正常发育或 当在疾病条件下被错误调节时，可能会使疱疹病毒核出口的研究成为 井。