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首页> 外文期刊>Nucleic Acids Research >The DEAD-box helicase Ded1 from yeast is an mRNP cap-associated protein that shuttles between the cytoplasm and nucleus
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The DEAD-box helicase Ded1 from yeast is an mRNP cap-associated protein that shuttles between the cytoplasm and nucleus

机译：来自酵母的DEAD-box解旋酶Ded1是一种mRNP帽相关蛋白，在细胞质和细胞核之间穿梭

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摘要

The DEAD-box helicase Ded1 is an essential yeast protein that is closely related to mammalian DDX3 and to other DEAD-box proteins involved in developmental and cell cycle regulation. Ded1 is considered to be a translation-initiation factor that helps the 40S ribosome scan the mRNA from the 5' 7-methylguanosine cap to the AUG start codon. We used IgG pull-down experiments, mass spectrometry analyses, genetic experiments, sucrose gradients, in situ localizations and enzymatic assays to show that Ded1 is a cap-associated protein that actively shuttles between the cytoplasm and the nucleus. NanoLC-MS/MS analyses of purified complexes show that Ded1 is present in both nuclear and cytoplasmic mRNPs. Ded1 physically interacts with purified components of the nuclear CBC and the cytoplasmic eIF4F complexes, and its enzymatic activity is stimulated by these factors. In addition, we show that Ded1 is genetically linked to these factors. Ded1 comigrates with these proteins on sucrose gradients, but treatment with rapamycin does not appreciably alter the distribution of Ded1; thus, most of the Ded1 is in stable mRNP complexes. We conclude that Ded1 is an mRNP cofactor of the cap complex that may function to remodel the different mRNPs and thereby regulate the expression of the mRNAs.
机译：DEAD-box解旋酶Ded1是必需的酵母蛋白，与哺乳动物DDX3和参与发育和细胞周期调控的其他DEAD-box蛋白密切相关。 Ded1被认为是一个翻译起始因子，可帮助40S核糖体扫描从5'7-甲基鸟苷帽到AUG起始密码子的mRNA。我们使用IgG下拉实验，质谱分析，遗传实验，蔗糖梯度，原位定位和酶促测定法来显示Ded1是与帽相关的蛋白，可在细胞质和细胞核之间有效地穿梭。纯化的复合物的NanoLC-MS / MS分析表明，Ded1存在于核和细胞质mRNP中。 Ded1与核CBC和细胞质eIF4F复合物的纯化成分发生物理相互作用，并且这些因子刺激其酶促活性。此外，我们表明Ded1与这些因素遗传相关。 Ded1在蔗糖梯度上与这些蛋白迁移，但是雷帕霉素处理并不会明显改变Ded1的分布。因此，大多数Ded1处于稳定的mRNP复合物中。我们得出的结论是，Ded1是帽复合物的mRNP辅因子，其功能可能是重构不同的mRNPs，从而调节mRNA的表达。

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