输入蛋白α可刺激帽甲基转移酶对GpppG - RNA的选择性结合和甲基化作用。
Cap methyltransferase selective binding and methylation of GpppG-RNA are stimulated by importin-alpha.
作者信息
Wen Y, Shatkin A J
机构信息
Center for Advanced Biotechnology and Medicine, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey 08854, USA.
出版信息
Genes Dev. 2000 Dec 1;14(23):2944-9. doi: 10.1101/gad.848200.
Abstract
We screened a human cDNA library for proteins that bind mRNA cap methyltransferase (MT) and isolated nuclear transporter importin-alpha (Impalpha). This direct association was confirmed by glutathione S-transferase (GST) pulldown, coimmunoprecipitation, and nuclear colocalization. In gel shift assays, MT selectively bound RNA containing 5'-terminal GpppG, and binding was inhibited by GpppG and not by m(7)GpppC. Impalpha markedly enhanced MT binding to GpppG-RNA and stimulated MT activity. MT/RNA/Impalpha complexes were dissociated by importin-beta, which also blocked the stimulation of cap methylation by Impalpha. The presence of RanGTP but not RanGDP prevented these effects of importin-beta. These findings indicate that importins play a novel role in mRNA biogenesis at the level of cap methylation.
摘要
我们在人cDNA文库中筛选与mRNA帽甲基转移酶（MT）结合的蛋白质，并分离出核转运蛋白输入蛋白α（Impα）。通过谷胱甘肽S-转移酶（GST）下拉实验、免疫共沉淀和细胞核共定位证实了这种直接关联。在凝胶迁移实验中，MT选择性结合含有5'-末端GpppG的RNA，且结合被GpppG抑制，而不被m(7)GpppC抑制。Impα显著增强MT与GpppG-RNA的结合并刺激MT活性。MT/RNA/Impα复合物被输入蛋白β解离，输入蛋白β也阻断了Impα对帽甲基化的刺激作用。RanGTP而非RanGDP的存在可阻止输入蛋白β的这些作用。这些发现表明输入蛋白在帽甲基化水平的mRNA生物合成中发挥新作用。
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