ABSTRACT 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：

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通过连续提取获得拟南芥综合核蛋白质组

摘要 在真核生物中，细胞核在基本细胞过程中起着关键作用，包括 DNA 复制、染色质维持、转录和翻译。为了更好地了解细胞核的功能多样性，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来纯化细胞核，然后进行基于溶解度的分馏以增加蛋白质组覆盖率。我们在拟南芥培养细胞的核部分中鉴定了 1539 种蛋白质和两种新型核膜 (NE) 蛋白质。通过 GFP 融合分析确定了 25 种蛋白质的定位；这些蛋白质中有 23 种位于细胞核或与 NE 相关的内质网中。该结果表明蛋白质组的高质量。