BRITE Pivot: Identification of the Biophysical Mechanisms Regulating Nuclear-Cytosolic Transport Capacity

这个促进工程变革和公平进步的研究思路（BRITE）枢纽研究项目的总体目标是全面了解大分子进出细胞核运动的生物物理调节。 细胞核是细胞中最大的结构，有两层膜包围着DNA。双膜有成千上万的小孔，称为核孔复合物（NPC）。NPC是分子（包括蛋白质和mRNA）进出细胞核的方式。这种分子通过NPC的运动是如此重要，哺乳动物（人类）和单细胞生物都有它。NPC运输的问题发生在许多疾病中，包括癌症，病毒感染，心血管疾病和衰老。 由于几乎每个细胞都有一个细胞核，因此测量分子在NPC中移动的新方法将有助于细胞生物学，生物物理学和机械生物学的研究。该项目的教育目标是通过在教育和推广经验中使用这些新的研究成果，向本科生介绍不断发展的核机械生物学领域。单个核孔复合物（NPC）的大分子结构已被非常详细地表征，包括许多核孔蛋白的定位和动力学。然而，NPC核运输的动力学和调节，在空间和时间层面上，还有待仔细研究，部分原因是在这些过程的检测工具的限制。在这个项目中，将开发和应用基于生物传感器的方法来更好地了解NPC核运输，以了解调节NPC运输的生化和生物力学因素。这些研究的结果将为尚未回答的关于NPC运输调节的基本问题提供实质性的见解：1）NPC运输活动是否存在空间差异？2)每个NPC都是专门进出口的吗？和3）机械负载的NPC改变核胞质转运？这项变革性的研究将为NPC核-胞质转运的生物物理调节提供新的理解。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The overall goal of this Boosting Research Ideas for Transformative and Equitable Advances in Engineering (BRITE) Pivot research project is to develop a comprehensive understanding of the biophysical regulation of the movement of large molecules in and out of the cell nucleus. The nucleus is the largest structure in a cell and has two membranes surrounding the DNA. The double membrane has thousands of small pores called nuclear pore complexes (NPCs). The NPCs are how molecules (including proteins and mRNA) move into and out of the nucleus. This movement of molecules through NPCs is so important that mammals (humans) and single-cell organisms all have it. Problems with NPC transport occurs in many diseases, including cancer, infection by viruses, cardiovascular disease, and aging. Because nearly every cell has a nucleus, the new methods for measuring the molecules moving through the NPCs will be useful for research in cell biology, biophysics, and mechanobiology. The educational goal of this project is to introduce undergraduate students to the growing field of nucleus mechanobiology by using these new research findings in educational and outreach experiences.The macromolecular structure of individual nuclear pore complexes (NPCs) has been characterized in great detail, including the localization and dynamics of many nucleoporin proteins. However, the dynamics and regulation of NPC nuclear transport, at a spatial and temporal level, have yet to be carefully studied, in part due to a limitation in tools for detection of these processes. In this project biosensor-based approaches will be developed and applied to better understand NPC nuclear transport in order to understand of the biochemical and biomechanical factors that regulate NPC transport. Results from these studies will provide substantial insight into yet-unanswered fundamental questions concerning NPC regulation of transport: 1) are there are spatial differences in NPC transport activity? 2) are individual NPCs specialized for import versus export? and 3) does mechanical loading of NPCs alter nuclear-cytosolic transport? This transformative research will provide a new understanding of the biophysical regulation of NPC nuclear-cytosolic transport.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.