Chemical Genetics Laboratory, RIKEN Advanced Science Institute, Wako, Saitama, 351-0198, Japan; Institute for Virus Research, Kyoto University, Sakyo-ku, Kyoto, 606-8507, Japan.
Genes Cells. 2014 Jan;19(1):78-87. doi: 10.1111/gtc.12114. Epub 2013 Dec 4.
To identify the novel factors involved in the postsplicing intron turnover pathway, we carried out immunoprecipitation with known postsplicing factors, hPrp43 and TFIP11. As an interacting factor, we identified C2ORF3 protein by mass spectrometry. We found that C2ORF3 protein is present in the previously characterized Intron Large (IL) complex with an excised lariat intron. In vitro splicing using C2ORF3-depleted nuclear extracts showed significant repression of splicing, suggesting that C2ORF3 protein is required for pre-mRNA splicing through its presumable role in efficient intron turnover. Interestingly, C2ORF3 protein is localized in both the nucleoplasm and nucleoli, which suggests a potential function in rRNA processing.
为了鉴定涉及剪接后内含子周转途径的新的因子,我们利用已知的剪接后因子 hPrp43 和 TFIP11 进行免疫沉淀。作为一个相互作用因子,我们通过质谱鉴定了 C2ORF3 蛋白。我们发现 C2ORF3 蛋白存在于以前表征的带有切除套索内含子的内含子大(IL)复合物中。使用耗尽 C2ORF3 的核提取物进行体外剪接显示剪接受到显著抑制,表明 C2ORF3 蛋白通过其在有效内含子周转中的假定作用而需要参与前体 mRNA 剪接。有趣的是,C2ORF3 蛋白定位于核质和核仁中,这表明其在 rRNA 加工中具有潜在功能。