Live imaging system for visualizing nuclear pore complex (NPC) formation during interphase in mammalian cells
Live imaging system for visualizing nuclear pore complex (NPC) formation during interphase in mammalian cells
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DOI：
10.1111/j.1365-2443.2010.01406.x
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发表时间：
2010-06-01
期刊：
影响因子：
2.1
通讯作者：
Imamoto, Naoko
中科院分区：
文献类型：
--
作者：
Iino, Haruki;Maeshima, Kazuhiro;Imamoto, Naoko
核孔复合体(NPC)是核膜上的超分子复合体，由大约30种不同蛋白质的多个拷贝组成，称为核孔蛋白(Nups)，它为间期核质运输提供水溶液通道。尽管鼻咽癌的结构特征已被详细描述，但鼻咽癌的形成及其调控，特别是在间期，还知之甚少。在这项研究中，利用温度敏感的RCC1突变体tsBN2，我们发现RCC1活性的缺乏抑制了间期NPC的形成，这表明在哺乳动物细胞间期NPC的形成需要RanGTP。利用tsBN2细胞中可逆的RCC1活性，我们建立了一个活细胞系统，允许通过温度变化来抑制或启动NPC的形成。我们的系统能够检测活细胞间期鼻咽癌的形成。由于RCC1的缺失减少了鼻咽癌结构中含有非结构化苯丙氨酸-甘氨酸重复序列的一些NUP，我们推测RCC1也参与了哺乳动物细胞间期维持NPC完整性的过程。
Nuclear pore complexes (NPCs) are 'supramolecular complexes' on the nuclear envelope assembled from multiple copies of approximately 30 different proteins called nucleoporins (Nups) that provide aqueous channels for nucleocytoplasmic transport during interphase. Although the structural aspects of NPCs have been characterized in detail, NPC formation and its regulation, especially during interphase, are poorly understood. In this study, using the temperature-sensitive RCC1 mutant tsBN2, a baby hamster kidney 21 cell line, we found that a lack of RCC1 activity inhibited NPC formation during interphase, suggesting that RanGTP is required for NPC formation during interphase in mammalian cells. Utilizing the reversible RCC1 activity in tsBN2 cells, we established a live-cell system that allows for the inhibition or initiation of NPC formation by changes in temperature. Our system enables the examination of NPC formation during interphase in living cells. As a lack of RCC1 decreased some Nups containing unstructured phenylalanine-glycine repeats in the NPC structure, we propose that RCC1 is also involved in maintaining NPC integrity during interphase in mammalian cells.