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Comprehensive nuclear proteome of Arabidopsis obtained by sequential extraction
Nucleus ( IF 3.7 ) Pub Date : 2019-04-09
Chieko Goto, Shoko Hashizume, Yoichiro Fukao, Ikuko Hara-Nishimura, Kentaro Tamura

In eukaryotes, the nucleus plays key roles in fundamental cellular processes, including DNA replication, chromatin maintenance, transcription, and translation. To better understand the functional diversity of nuclei, we developed a method for the comprehensive extraction of the nuclear proteome from Arabidopsis. We used a buffer with a high sucrose concentration to purify nuclei and then conducted solubility-based fractionation to increase proteome coverage. We identified 1539 proteins and two novel nuclear envelope (NE) proteins in the nuclear fraction of Arabidopsis cultured cells. The localization of 25 proteins was determined by GFP fusion analyses; 23 of these proteins were localized either in the nucleus or the NE-associated endoplasmic reticulum. This result was indicative of the high quality of the proteome. These findings will be useful for clarifying novel nuclear functions in plants.



中文翻译:

通过顺序提取获得的拟南芥综合核蛋白质组

在真核生物中,核在基本的细胞过程中起关键作用,包括DNA复制,染色质维持,转录和翻译。为了更好地了解核的功能多样性,我们开发了一种从拟南芥中全面提取核蛋白质组的方法。我们使用具有高蔗糖浓度的缓冲液纯化细胞核,然后进行基于溶解度的分级分离以增加蛋白质组覆盖率。我们在拟南芥的核级分中鉴定了1539个蛋白和两个新的核包膜(NE)蛋白培养的细胞。通过GFP融合分析确定了25种蛋白质的定位。这些蛋白质中的23个位于核或与NE相关的内质网中。该结果表明蛋白质组的高质量。这些发现将有助于阐明植物中新的核功能。

更新日期:2019-04-09
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