项目名称： 三维超分辨技术研究细胞核孔复合物的选择性屏障分布

项目编号： No.31500599

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 马炯

作者单位： 复旦大学

项目金额： 21万元

中文摘要： 细胞核孔复合物（NPC）是真核细胞中连接细胞质和细胞核之间的选择性双向通道。由富含苯丙氨酸-甘氨酸（FG）氨基酸片段的自然非折叠蛋白构成的FG-屏障,选择性地调控基因相关物质,按照被动运输或者主动介入运输的方式进出细胞核。然而，核孔内FG屏障间的相互作用以及分布状况仍然是极具争议的重要科学问题。本项目将基于申请人发明的单点斜入射激发快速超分辨显微镜技术，研究核孔传输蛋白或FG-蛋白片段与FG屏障反应的三维分布图像，从而获得FG-屏障在细胞核孔内完整的分布情况。另外,进一步检测核孔传输蛋白对FG屏障的分布影响，讨论核孔传输的自调控机制。还将从不同传输蛋白或FG-片段在传输过程中的竞争效果，讨论FG屏障对各类基因相关物质传输的优先顺序。最终，完整的三维FG屏障将对核孔蛋白缺失相关的白血病、阿尔兹海默症的致病机制，以及各种类病毒的侵入机制等核孔相关病理机制或基因治疗提供信息。

中文关键词： 细胞核孔复合物；FG屏障；单分子；超分辨显微镜；核孔传输

英文摘要： The nuclear pore complexes (NPCs) selectively mediate the bidirectional trafficking of macromolecules between the cytoplasm and the nucleus in eukaryotic cells. The selective barrier formed by natively unfolded phenylalanine-glycine (FG) nucleoporins (Nups) inside the NPC allows for passive and facilitated transport through the NPC. However, the interactions that form the FG-barrier and their spatial distribution in the NPC remain unresolved. Based on the single-point edge-excitation sub-diffusion (SPEED) microscopy techniques, the applicant's invention, three-dimensionally map the interaction distributions between the fluorescent transport receptors (TRs) or FG-fragment and FG barrier. Then, we obtain fully information of the FG-barrier distribution in the NPC. The conformation change of the FG-barrier by TRs would be discussed as self-regulating mechanism of nucleocytoplasmic transport. Also from the competitive effects during different FG-fragments and transport proteins during the transport discuss priorities for the various gene related transport. Finally，the complete three-dimensional FG-barrier distribution will provide information with leukemia, Alzheimer's disease pathogenesis, as well as various types of viruses invade mechanisms for NPC related pathological mechanism or gene therapy.

英文关键词： Nuclear pore complex;FG barrier;single-molecule;Super-resolution microscopy;Nucleocytoplasmic transport