Investigating nucleo-cytoplasmic partitioning in Alzheimer's disease and aging

项目摘要/摘要 蛋白质的核质分配是一种新兴的细胞内过程，具有至关重要的作用 在阿尔茨海默氏症和衰老方面。核粘附素通过转运介导适当的分配 蛋白质穿过核孔。核粘附素的药理调节提供了一种新的 治疗阿尔茨海默病的方法。 最近对模式生物和哺乳动物细胞的研究发现，有几种 神经退行性疾病表现为核浆蛋白分裂功能障碍。我们 有报道称，保守的核粘附素XPO1通过促进核老化而加速衰老 输出与长寿相关的转录因子，从而阻止维持 蛋白质抑制机制。具体地说，我们证明了抑制XPO1可以延长寿命 延伸并导致整个门的蛋白平衡增强，部分通过自噬- 溶酶体途径。长寿动物的XPO1水平降低，我们最近的数据 提示XPO1介导的寿命调节依赖于转录组的根本变化 和蛋白质组的分割以及核仁的重组，这导致核糖体的改变 生物发生学。因此，我们假设XPO1协调调节核质 蛋白质的分配和核仁的形成，从而调节隔室特有的蛋白质 功能，核糖体的生物发生，活性信使核糖核酸的翻译，从而寿命。 为了解决这一假设，我们建议对蛋白质进行全球表征 分割和规范及其对蛋白质代谢和衰老的影响。我们将结合切割- 线虫和哺乳动物细胞的边缘遗传学和生化方法 衰老过程中蛋白质的核质分配(目标1)，并最终发现 新的核仁功能调节剂具有潜在的预防阿尔茨海默病的益处(目标2)。 这些拟议的研究具有重要意义，因为它们解决了与衰老相关的一个重要问题 核质分配和核仁功能在生物体中的作用和调节 衰老。我们的方法是创新的，因为我们并行利用线虫和哺乳动物细胞 快速识别在衰老中起作用的特定蛋白质的翻译和分配，并 发现调节核仁和核糖体动力学的药理策略 预防阿尔茨海默氏症。

PROJECT SUMMARY/ABSTRACT Nucleo-cytoplasmic partitioning of proteins is an emerging intracellular process with crucial roles in Alzheimer’s disease and aging. Karyopherins mediate proper partitioning by transporting proteins across the nuclear pore. Pharmacological modulation of karyopherins provides a new approach for therapies against Alzheimer’s disease. Recent studies in model organisms and mammalian cells have uncovered that several neurodegenerative diseases display dysfunctional nucleo-cytoplasmic protein partitioning. We have reported that the conserved karyopherin XPO1 accelerates aging by promoting the nuclear export of longevity-associated transcription factors, thereby preventing the maintenance of proteostatic mechanisms. Specifically, we showed that inhibiting XPO1 leads to lifespan extensions and results in enhanced proteostasis across phyla, in part via the autophagy- lysosomal pathway. Levels of XPO1 are reduced in long-lived animals and our recent data suggest that XPO1-mediated lifespan modulation relies on fundamental changes in transcriptome and proteome partitioning as well as nucleoli re-organization, which results in altered ribosomal biogenesis. Thus, we hypothesize that XPO1 coordinately modulates the nucleo-cytoplasmic partitioning of proteins and nucleoli formation, thereby regulating compartment-specific protein functions, ribosomal biogenesis, active mRNA translation and consequently lifespan. To address this hypothesis, we propose to conduct a global characterization of protein partitioning and specification and its impact on proteostasis and aging. We will combine cutting- edge genetics and biochemical approaches in C. elegans and mammalian cells to characterize the nucleo-cytoplasmic partitioning of proteins during aging (Aim 1) and, ultimately, to uncover new modulators of nucleolar function with potential benefits against Alzheimer’s disease (Aim 2). These proposed studies are significant as they address an important question in aging related to the role and regulation of nucleo-cytoplasmic partitioning and nucleoli function during organism aging. Our approaches are innovative since we utilize C. elegans and mammalian cells in parallel to quickly identify specific protein translation and partitioning of proteins with roles in aging and to uncover pharmacological strategies to modulate nucleolar and ribosomal dynamics and preventing Alzheimer’s disease.