Directed proteomic analysis of the human nucleolus
Directed proteomic analysis of the human nucleolus
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DOI：
10.1016/s0960-9822(01)00650-9
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发表时间：
2002-01-08
期刊：
影响因子：
9.2
通讯作者：
Lamond, AI
中科院分区：
文献类型：
--
作者：
Andersen, JS;Lyon, CE;Lamond, AI
背景：核仁是含有核糖体RNA基因簇和核糖体生物发生因子的亚核细胞器。最近的研究表明，它还可能在RNA运输、RNA修饰和细胞周期调节中发挥作用。尽管对核仁进行了150多年的研究，但其结构和功能的许多方面仍不清楚。结果：我们报道了对人类核仁的蛋白质组学分析。使用质谱仪(MS)和序列数据库搜索的组合，包括对起草的人类基因组序列的在线分析，鉴定出271种蛋白质。超过30%的核仁蛋白是由新的或未知的基因编码的，而已知的蛋白包括一些未知的因子，这些因子是以前没有已知的核仁功能的。从转录被抑制的HeLa细胞中分离出的核仁的MS分析表明，蛋白质的一个子集被浓缩。这些数据突出了核仁蛋白质组的动态性质，并表明蛋白质可以与核仁短暂结合，也可以仅在特定的代谢条件下积累。结论：这种广泛的蛋白质组学分析表明，核仁具有惊人的蛋白质复杂性。已确定的许多新的因子和不同类别的蛋白质支持这样的观点，即核仁可能在核糖体亚单位生物发生中发挥其已知作用之外的额外功能。数据还表明，核仁的蛋白质组成并不是一成不变的，它可以随着细胞的代谢状态而发生显著变化。背景：核仁是一种亚核细胞器，包含核糖体RNA基因簇和核糖体生物发生因子。最近的研究表明，它还可能在RNA运输、RNA修饰和细胞周期调节中发挥作用。尽管对核仁进行了150多年的研究，但其结构和功能的许多方面仍不清楚。结果：我们报道了对人类核仁的蛋白质组学分析。使用质谱仪(MS)和序列数据库搜索的组合，包括对起草的人类基因组序列的在线分析，鉴定出271种蛋白质。超过30%的核仁蛋白是由新的或未知的基因编码的，而已知的蛋白包括一些未知的因子，这些因子是以前没有已知的核仁功能的。HeLa核仁的MS分析转录被抑制的细胞表明，蛋白质的一个子集得到了浓缩。这些数据突出了核仁蛋白质组的动态性质，并表明蛋白质可以与核仁短暂结合，也可以仅在特定的代谢条件下积累。结论：这种广泛的蛋白质组学分析表明，核仁具有惊人的蛋白质复杂性。已确定的许多新的因子和不同类别的蛋白质支持这样的观点，即核仁可能在核糖体亚单位生物发生中发挥其已知作用之外的额外功能。数据还表明，核仁的蛋白质组成并不是一成不变的，可以随着细胞的新陈代谢状态而发生显著变化。
Background: The nucleolus is a subnuclear organelle containing the ribosomal RNA gene clusters and ribosome biogenesis factors. Recent studies suggest it may also have roles in RNA transport, RNA modification, and cell cycle regulation. Despite over 150 years of research into nucleoli, many aspects of their structure and function remain uncharacterized.Results: We report a proteomic analysis of human nucleoli. Using a combination of mass spectrometry (MS) and sequence database searches, including online analysis of the draft human genome sequence, 271 proteins were identified. Over 30% of the nucleolar proteins were encoded by novel or uncharacterized genes, while the known proteins included several unexpected factors with no previously known nucleolar functions. MS analysis of nucleoli isolated from HeLa cells in which transcription had been inhibited showed that a subset of proteins was enriched. These data highlight the dynamic nature of the nucleolar proteome and show that proteins can either associate with nucleoli transiently or accumulate only under specific metabolic conditions.Conclusions: This extensive proteomic analysis shows that nucleoli have a surprisingly large protein complexity. The many novel factors and separate classes of proteins identified support the view that the nucleolus may perform additional functions beyond its known role in ribosome subunit biogenesis. The data also show that the protein composition of nucleoli is not static and can alter significantly in response to the metabolic state of the cell.Background: The nucleolus is a subnuclear organelle containing the ribosomal RNA gene clusters and ribosome biogenesis factors. Recent studies suggest it may also have roles in RNA transport, RNA modification, and cell cycle regulation. Despite over 150 years of research into nucleoli, many aspects of their structure and function remain uncharacterized.Results: We report a proteomic analysis of human nucleoli. Using a combination of mass spectrometry (MS) and sequence database searches, including online analysis of the draft human genome sequence, 271 proteins were identified. Over 30% of the nucleolar proteins were encoded by novel or uncharacterized genes, while the known proteins included several unexpected factors with no previously known nucleolar functions. MS analysis of nucleoli isolated from HeLa. cells in which transcription had been inhibited showed that a subset of proteins was enriched. These data highlight the dynamic nature of the nucleolar proteome and show that proteins can either associate with nucleoli transiently or accumulate only under specific metabolic conditions.Conclusions: This extensive proteomic analysis shows that nucleoli have a surprisingly large protein complexity. The many novel factors and separate classes of proteins identified support the view that the nucleolus may perform additional functions beyond its known role in ribosome subunit biogenesis. The data also show that the protein composition of nucleoli is not static and can alter significantly in response to the metabolic state of the cell.