Nuclear dynamics in Arabidopsis thaliana
Nuclear dynamics in Arabidopsis thaliana
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DOI：
10.1091/mbc.11.8.2733
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发表时间：
2000-08-01
影响因子：
3.3
通讯作者：
Galbraith, DW
中科院分区：
文献类型：
--
作者：
Chytilova, E;Macas, J;Galbraith, DW
细胞核是真核细胞的决定性特征，其包括双双层膜，即内核膜和外核膜，其将核质和细胞质隔室分开。核孔，连接两个膜的复杂大分子组件，介导这些隔室之间的通信。为了探索活植物细胞内细胞核的形态、拓扑结构和动力学，我们开发了一种在延时条件下共聚焦激光扫描荧光显微镜的新方法。这用于检测嵌合蛋白在拟南芥中的转基因表达，所述嵌合蛋白包含与有效的核定位信号（NLS）和来自E.杆菌这种大蛋白质靶向细胞核并仅在核质内积累。本文提供了在线访问电影，这些电影说明了细胞核显示的显着和不寻常的特性，包括多态形状变化和快速，长距离，细胞内运动。运动是由肌动蛋白介导的，但不是由微管蛋白，因此，它似乎不同于核定位和迁移的机制，已报告的真核生物。基于GFP的检测方法简单，具有普遍适用性。这将是有趣的，以确定是否在其他真核生物中观察到的新类型的动态行为，在这里报告的高等植物。
The nucleus is a definitive feature of eukaryotic cells, comprising twin bilamellar membranes, the inner and outer nuclear membranes, which separate the nucleoplasmic and cytoplasmic compartments. Nuclear pores, complex macromolecular assemblies that connect the two membranes, mediate communication between these compartments. To explore the morphology, topology, and dynamics of nuclei within living plant cells, we have developed a novel method of confocal laser scanning fluorescence microscopy under time-lapse conditions. This is used for the examination of the transgenic expression in Arabidopsis thaliana of a chimeric protein, comprising the GFP (Green-Fluorescent Protein of Aequorea victoria) translationally fused to an effective nuclear localization signal (NLS) and to beta-glucuronidase (GUS) from E. coli. This large protein is targeted to the nucleus and accumulates exclusively within the nucleoplasm.This article provides online access to movies that illustrate the remarkable and unusual properties displayed by the nuclei, including polymorphic shape changes and rapid, long-distance, intracellular movement. Movement is mediated by actin but not by tubulin; it therefore appears distinct from mechanisms of nuclear positioning and migration that have been reported for eukaryotes. The GFP-based assay is simple and of general applicability. It will be interesting to establish whether the novel type of dynamic behavior reported here, for higher plants, is observed in other eukaryotic organisms.