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.

项目摘要/摘要 （如父母赠款所述） 核酸和蛋白质从核从细胞质到细胞质的转运几乎是Ally Allularr 过程以及误认为的过程与疾病，肿瘤形成/生长和癌症有关 在规范上，这种必不可少的过程被认为是通过核 孔复合体，沿核心包膜的双膜横跨孔复合物，并提供关键的常规 步骤INWAT退出（并进入）核，核心包膜（Ne-）的萌芽被证明 为核出口提供了替代途径，特别是针对大型核糖核蛋白（RNP） 否则，需要展开/改建以适合毛孔。 大分子复合物被内核膜封装，越过核周 空间，与外核膜融合，并释放到细胞质中，一种机制 与疱疹病毒核的出口非常相似。 复合物一起退出核，并作为特定细胞功能的包装磨碎。 尽管具有明显的生物学重要性和临床相关性，但对调节的知之甚少 或最近在任何系统中发生的结构机械。 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对于使用WASH/SHRC作为新的入口点是必需的 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。 了解政府的分子和细胞力学。 建议是确定Ne-Budding中WASH/SHRC功能的机制，并 识别/分析控制动态NE-BUDDING过程的基础结构组件/机械 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。 用于研究此保守过程的生物是成像和切割的财富的能力 边缘/分子技术和试剂。 阐明在正常发育或 当在疾病条件下进行误导时，并且可以告知疱疹病毒核心的研究 出色地。