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: 10541746
• 负责人: SUSAN M PARKHURST
• 金额: $ 8.55万
• 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/10541746
• 关键词: Actins; Aging; Biochemical; Biological; Cell Nucleus; Cell physiology; Cells; Clinical; Complex; Cytoplasm; Development; Developmental Process; Disease; Drosophila genus; Due Process; Encapsulated; Family; Gene Proteins; Genetic; Goals; Growth; Herpesviridae; Herpesviridae Infections; Image; Immune; Infrastructure; Malignant Neoplasms; Mechanics; Membrane; Microscopy; Molecular; Nuclear; Nuclear Envelope; Nuclear Export; Nuclear Inner Membrane; Nuclear Outer Membrane; Nuclear Pore Complex; Nucleic Acids; Organism; Pathway interactions; Process; Proteins; Reagent; Regulation; Resolution; Ribonucleoproteins; Symptoms; System; Techniques; Time; Transport Process; Wiskott-Aldrich Syndrome; clinically relevant; insight; macromolecule; nervous system disorder; novel; parent grant; therapeutic development; tumor; tumor progression

PROJECT SUMMARY/ABSTRACT (as stated in parent grant) 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.

项目概要/摘要 （如家长补助金中所述） 核酸和蛋白质从细胞核到细胞质的运输对于几乎所有细胞都至关重要 当调节不当时，这些过程与疾病、肿瘤形成/生长和癌症有关 进展。按照规范，这一不可或缺的过程被认为仅通过核发生 孔复合物，跨越核膜的双层膜并提供关键的调节 进入（和进入）细胞核的步骤。最近，核膜（NE-）出芽被证明可以 提供核退出的替代途径，特别是对于大核糖核蛋白（RNP）复合物 否则需要展开/重塑以适应毛孔。在这条道路上，大 大分子复合物被内核膜包裹，穿过核周 空间，与外核膜融合，并释放到细胞质中，这是一种机制 与疱疹病毒核排出惊人相似。因此，NE 出芽优雅地允许大 RNP 复合物一起离开细胞核并作为特定细胞功能的包进行递送。 尽管其具有明显的生物学重要性和临床相关性，但人们对其监管知之甚少。 或允许在任何系统中发生 NE 出芽的结构机械。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 出芽是必需的。串联使用 WASH/SHRC 作为新的切入点 通过发现参与该过程的新基因/蛋白质的策略，我们的长期目标是 了解控制 NE 出芽的分子和细胞力学。本次活动的具体目标 建议确定 WASH/SHRC 在 NE 萌芽中的功能机制，并 识别/分析控制动态 NE 萌芽过程的基础设施组件/机械 结合遗传、生化、细胞生物学、延时实时成像和超级 分辨率/电磁显微镜方法。果蝇提供了一种优秀的、遗传上顺从的、 由于其易于成像和丰富的切割能力，因此成为研究这一保守过程的生物体 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 阐明在正常发展或 当在疾病条件下受到错误调节时，可能会为疱疹病毒核排出的研究提供信息 出色地。