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首页> 外文期刊>Nucleus >Characterization of nuclear pore complex components in fission yeast Schizosaccharomyces pombe
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Characterization of nuclear pore complex components in fission yeast Schizosaccharomyces pombe

机译：裂殖酵母裂殖酵母核孔复合体成分的表征

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摘要

The nuclear pore complex (NPC) is an enormous proteinaceous complex composed of multiple copies of about 30 different proteins called nucleoporins. In this study, we analyzed the composition of the NPC in the model organism Schizosoccharomyces pombe using strains in which individual nucleoporins were tagged with GFP. We identified 31 proteins as nucleoporins by their localization to the nuclear periphery. Gene disruption analysis in previous studies coupled with gene disruption analysis in the present study indicates that 15 of these nucleoporins are essential for vegetative cell growth and the other 16 nucleoporins are non-essential. Among the 16 non-essential nucleoporins, 11 are required for normal progression through meiosis and their disruption caused abnormal spore formation or poor spore viability. Based on fluorescence measurements of GFP-fused nucleoporins, we estimated the composition of the NPC in S. pombe and found that the organization of the S. pombe NPC is largely similar to that of other organisms; a single NPC was estimated as being 45.8-47.8 MDa in size. We also used fluorescence measurements of single NPCs and quantitative western blotting to analyze the composition of the Nup107-Nup160 subcomplex, which plays an indispensable role in NPC organization and function. Our analysis revealed low amounts of Nup107 and Nup131 and high amounts of Nup132 in the Nup107-Nup160 subcomplex, suggesting that the composition of this complex in S. pombe may differ from that in S. cerevisiae and humans. Comparative analysis of NPCs in various organisms will lead to a comprehensive understanding of the functional architecture of the NPC.
机译：核孔复合物（NPC）是一种巨大的蛋白质复合物，由大约30种称为核孔蛋白的不同蛋白质的多个副本组成。在这项研究中，我们分析了模型生物体粟酒裂殖酵母中NPC的组成，该菌株使用了单个核孔蛋白标记有GFP的菌株。我们通过定位到核外围将31种蛋白鉴定为核孔蛋白。先前研究中的基因破坏分析以及本研究中的基因破坏分析表明，这些核孔蛋白中有15种对于营养细胞的生长是必不可少的，而其他16种核孔蛋白则不是必需的。在16种非必需的核孔蛋白中，通过减数分裂的正常进程需要11种，而它们的破坏会导致异常的孢子形成或不良的孢子生存能力。根据GFP融合核孔蛋白的荧光测量结果，我们估算了粟酒裂殖酵母中NPC的组成，发现粟酒裂殖酵母NPC的组织与其他生物体非常相似。单个NPC的大小估计为45.8-47.8 MDa。我们还使用了单个NPC的荧光测量和定量Western印迹分析了Nup107-Nup160亚复合物的组成，该复合物在NPC的组织和功能中起着不可或缺的作用。我们的分析显示，Nup107-Nup160亚复合物中Nup107和Nup131的含量较低，Nup132的含量较高，这表明粟酒裂殖酵母中该复合物的成分可能与酿酒酵母和人中的有所不同。对各种生物中的NPC进行比较分析将使人们对NPC的功能结构有一个全面的了解。

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