Hirano Yasuhiro, Ishii Kohei, Kumeta Masahiro, Furukawa Kazuhiro, Takeyasu Kunio, Horigome Tsuneyoshi
Kyoto University, Yoshida-konoe-cho, Kyoto, Japan.
Genes Cells. 2009 Feb;14(2):155-66. doi: 10.1111/j.1365-2443.2008.01262.x. Epub 2008 Jan 12.
The nuclear matrix has classically been assumed to be a solid structure coherently aligning nuclear components, but its real nature remains obscure. We separated the proteins in a ribonucleoprotein-containing nuclear matrix fraction of HeLa cells by reversed-phase HPLC followed by SDS-PAGE, and identified 83 proteins through peptide mass fingerprint (PMF) analysis. Many nucleolar proteins, classical nuclear matrix proteins, RNA binding proteins, cytoskeletal proteins and five uncharacterized proteins were identified in this fraction. Four of the latter proteins were localized to the cell nucleus, BXDC1 and EBNA1BP2 being especially localized to the nucleolus. Fluorescence recovery after photobleaching and RNAi knockdown analyses suggested that BXDC1 and EBNA1BP2 function in a dynamic scaffold for ribosome biogenesis.
传统上认为核基质是一种使核成分连贯排列的固体结构，但其真实性质仍不清楚。我们通过反相高效液相色谱（HPLC）随后进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE），分离了HeLa细胞含核糖核蛋白的核基质部分中的蛋白质，并通过肽质量指纹（PMF）分析鉴定了83种蛋白质。在该部分中鉴定出许多核仁蛋白、经典核基质蛋白、RNA结合蛋白、细胞骨架蛋白和五种未鉴定的蛋白质。后四种蛋白质中有四种定位于细胞核，BXDC1和EBNA1BP2尤其定位于核仁。光漂白后的荧光恢复和RNA干扰敲低分析表明，BXDC1和EBNA1BP2在核糖体生物发生的动态支架中发挥作用。
