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Three-Dimensional Mapping of mRNA Export through the Nuclear Pore Complex

机译:通过核孔复合体mRNA出口的三维映射。

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The locations of transcription and translation of mRNA in eukaryotic cells are spatially separated by the nuclear envelope (NE). Plenty of nuclear pore complexes (NPCs) embedded in the NE function as the major gateway for the export of transcribed mRNAs from the nucleus to the cytoplasm. Whereas the NPC, perhaps one of the largest protein complexes, provides a relatively large channel for macromolecules to selectively pass through it in inherently three-dimensional (3D) movements, this channel is nonetheless below the diffraction limit of conventional light microscopy. A full understanding of the mRNA export mechanism urgently requires real-time mapping of the 3D dynamics of mRNA in the NPC of live cells with innovative imaging techniques breaking the diffraction limit of conventional light microscopy. Recently, super-resolution fluorescence microscopy and single-particle tracking (SPT) techniques have been applied to the study of nuclear export of mRNA in live cells. In this review, we emphasize the necessity of 3D mapping techniques in the study of mRNA export, briefly summarize the feasibility of current 3D imaging approaches, and highlight the new features of mRNA nuclear export elucidated with a newly developed 3D imaging approach combining SPT-based super-resolution imaging and 2D-to-3D deconvolution algorithms.
机译:真核细胞中mRNA转录和翻译的位置在空间上被核被膜(NE)隔开。 NE中嵌入的大量核孔复合物(NPC)充当了转录的mRNA从细胞核到细胞质输出的主要通道。尽管NPC可能是最大的蛋白质复合物之一,但它为大分子提供了一个相对较大的通道,使其以固有的三维(3D)运动方式有选择地通过它,但该通道仍低于常规光学显微镜的衍射极限。迫切需要对mRNA出口机制的全面了解,并通过创新的成像技术实时绘制活细胞NPC中mRNA的3D动态图,从而打破传统光学显微镜的衍射极限。最近,超分辨率荧光显微镜和单粒子跟踪(SPT)技术已被用于研究活细胞中mRNA的核输出。在这篇综述中,我们强调在mRNA出口研究中使用3D映射技术的必要性,简要总结了当前3D成像方法的可行性,并着重介绍了新开发的结合基于SPT的3D成像方法阐明的mRNA核输出的新特征超分辨率成像和2D到3D反卷积算法。

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