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: 10642008
• 负责人: SUSAN M PARKHURST
• 金额: $ 17.74万
• 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/10642008
• 关键词: Cellular; mechanisms; nucleocytoplasmic; export; through

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.

项目总结/摘要 核酸和蛋白质从细胞核到细胞质的运输对于几乎所有的细胞生物学都是必需的。 过程，并且当被错误调节时，与疾病、肿瘤形成/生长和癌症相关。 进展规范上，这一不可或缺的过程被认为是完全通过核发生的。 孔复合物，它跨越核膜的双层膜，并提供关键的调节 在什么步骤退出（和进入）的核心。最近，核膜（NE-）出芽被证明是 为核出口提供替代途径，特别是对于大的核糖核蛋白（RNP）复合物 否则将需要展开/重塑以适合通过孔隙。在这条道路上，大 大分子复合物被内核膜包裹，穿过核周膜， 空间，与外核膜融合，并被释放到细胞质中，这是一种机制， 与疱疹病毒的核扩散惊人相似因此，NE-出芽优雅地允许大的RNP 复合物一起离开细胞核，并作为特定细胞功能的包被递送。 尽管其具有明确的生物学重要性和临床相关性，但关于其调控机制仍知之甚少。 或结构机制，使NE-萌芽发生在任何系统。最近，我们发现， Wiskott Aldrich综合征家族肌动蛋白成核蛋白，WASH，其四亚基调节复合物 （SHRC）和Arp 2/3是NE芽形成所必需的。将水、环境卫生和个人卫生以及健康和生殖健康中心作为新的切入点， 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NE-出芽的分子和细胞机制。具体目标是 建议是确定WASH/SHRC在NE-萌芽中的功能机制，并 确定/分析控制动态东北萌芽过程的基础设施组件/机制 利用遗传学、生物化学、细胞生物学、延时实时成像和超 分辨率/EM显微镜方法。果蝇提供了一个优秀的，遗传上顺从的， 由于其易于成像和丰富的切割，因此是研究这一保守过程的有机体 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 阐明这种令人兴奋的新核出口途径在正常发展中的机制， 当在疾病条件下错误调节时，并且可以告知疱疹病毒核出口的研究， 好.