聚腺苷酸结合蛋白对信使核糖核酸的稳定作用不依赖于聚腺苷酸且需要翻译过程。
mRNA stabilization by poly(A) binding protein is independent of poly(A) and requires translation.
作者信息
Coller J M, Gray N K, Wickens M P
机构信息
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706-1544 USA.
出版信息
Genes Dev. 1998 Oct 15;12(20):3226-35. doi: 10.1101/gad.12.20.3226.
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 we describe may be useful in identifying and assaying 3' UTR regulatory proteins, as it uncouples analysis of function from RNA binding.
多聚腺苷酸（poly(A)）尾的存在可增强翻译和mRNA稳定性。在体内，该尾与一种保守的多肽——多聚腺苷酸结合蛋白（Pab1p）相互作用。为了在体内检测Pab1p的功能，我们通过将Pab1p与MS2外壳蛋白融合，并在报告基因mRNA的3'非翻译区（UTR）中放置MS2结合位点，从而将Pab1p连接到报告基因mRNA的3'UTR上。这种策略使我们能够将Pab1p的功能与其RNA结合活性分离。我们发现，体内缺乏多聚腺苷酸尾的mRNA可被Pab1p稳定，并且稳定所需的Pab1p部分在基因上与多聚腺苷酸结合所需的部分不同。此外，Pab1p介导的稳定作用需要mRNA持续翻译。我们得出结论，就mRNA稳定性而言，多聚腺苷酸的主要或唯一功能仅仅是将Pab1p带到mRNA上，并且mRNA稳定是Pab1p的固有特性。我们描述的方法可能有助于鉴定和分析3'UTR调控蛋白，因为它将功能分析与RNA结合分离。
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