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转移到细胞质，对于几乎所有细胞都是必不可少的 过程以及误导的过程与疾病，肿瘤形成/生长和癌症有关 进展。从规范上讲，这一必不可少的过程被认为仅通过核发生 毛孔复合物，沿核包膜的双膜膜并提供关键的调节 介入出口（并进入）核的东西。最近，核包络（NE-）发芽显示为 提供核出口的替代途径，特别是对于大核糖核蛋白（RNP）配合物 否则，这将需要展开/改建以通过毛孔进行安装。在这条路中，很大 大分子复合物被内部核膜封装，越过核周 空间，与外核膜融合，并释放到细胞质中，这是一种机制 与疱疹病毒核出口非常相似。这是优雅的budding允许大型RNP 复合物一起退出核并作为特定细胞功能的包装交付。 尽管具有明显的生物学重要性和临床相关性，但对调节的知识仍然很少 或结构机器，可以在任何系统中发生NE-BUDDING。最近，我们发现 Wiskott Aldrich综合征家族肌动蛋白成核蛋白，WASH，其四个亚基调节复合物 （SHRC）和ARP2/3对于Ne-Budding是必需的。使用WASH/SHRC作为新的入口点 通过发现这一过程中涉及的新基因/蛋白质的策略，我们的长期目标是 了解控制NE-Budding的分子和细胞力学。这个特定的目的 建议是确定Ne-Budding中WASH/SHRC功能的机制，并 识别/分析管理动态NE-BUDDING过程的基础设施组件/机器 结合遗传，生化，细胞生物学，延时活成像和超级 分辨率/EM显微镜方法。果蝇提供了一种出色的，遗传上的良好， 有机体用于研究这种保守的过程，因为它的成像和切割丰富的财富 边缘细胞/分子技术和试剂。这些研究中收集的信息将有助于 阐明在正常发育或 当在疾病条件下误导并可能将疱疹病毒核出口的研究告知 出色地。