mRNA stabilization by poly(A) binding protein is independent of poly(A) and requires translation
mRNA stabilization by poly(A) binding protein is independent of poly(A) and requires translation
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DOI：
10.1101/gad.12.20.3226
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发表时间：
1998-10-15
影响因子：
10.5
通讯作者：
Wickens, MP
中科院分区：
文献类型：
--
作者：
Coller, JM;Gray, NK;Wickens, MP
翻译和mRNA稳定性通过poly（A）尾的存在而增强。在体内，尾部与保守多肽poly（A）结合蛋白（Pab1p）相互作用。为了检测Pab1p在体内的功能，我们通过将Pab1p与MS2外壳蛋白融合并将MS2结合位点置于报告基因的3'UTR中，将Pab1p与报告基因mRNA的3'UTR连接。这种策略使我们能够将Pab1p的功能与其RNA结合活性解偶联。我们表明，在体内缺乏poly（A）尾的mRNA被Pab1p稳定，并且稳定所需的Pab1p部分在遗传上与poly（A）结合所需的部分不同。此外，Pab1p的稳定需要mRNA的持续翻译。我们的结论是，主要的，或唯一的，功能的poly（A）相对于mRNA的稳定性是简单地把Pab1p的mRNA，和mRNA的稳定性是Pab1p的内在属性。rye描述的方法可用于鉴定和测定3'UTR调节蛋白，因为它将功能分析与RNA结合分离。
Translation and mRNA stability are enhanced by the presence of a poly(A) tail. In vivo, the tail interacts with a conserved polypeptide, poly(A) binding protein (Pab1p). To examine Pab1p function in vivo, we have tethered Pab1p to the 3' UTR of reporter mRNAs by fusing it to MS2 coat protein and placing MS2 binding sites in the 3' UTR of the reporter. This strategy allows us to uncouple Pab1p function from its RNA binding activity. We show that mRNAs that lack a poly(A) tail in vivo are stabilized by Pab1p, and that the portions of Pab1p required for stabilization are genetically distinct from those required for poly(A) binding. In addition, stabilization by Pab1p requires ongoing translation of the mRNA. We conclude that the primary, or sole, function of poly(A) with respect to mRNA stability is simply to bring Pab1p to the mRNA, and that mRNA stabilization is an intrinsic property of Pab1p. The approach rye describe may be useful in identifying and assaying 3' UTR regulatory proteins, as it uncouples analysis of function from RNA binding.