From the Departments of Molecular and Cellular Biochemistry and.
J Biol Chem. 2014 Feb 7;289(6):3703-12. doi: 10.1074/jbc.M113.538264. Epub 2013 Dec 20.
Insufficient expression of the survival motor neuron (SMN) protein causes spinal muscular atrophy, a neurodegenerative disease characterized by loss of motor neurons. Despite the importance of maintaining adequate SMN levels, little is known about factors that control SMN expression, particularly 3' end processing of the SMN pre-mRNA. In this study, we identify the U1A protein as a key regulator of SMN expression. U1A, a component of the U1 snRNP, is known to inhibit polyadenylation upon direct binding to mRNA. We show that U1A binds directly and with high affinity and specificity to the SMN 3'-UTR adjacent to the polyadenylation site, independent of the U1 snRNP (U1 small nuclear ribonucleoprotein). Binding of U1A inhibits polyadenylation of the SMN pre-mRNA by specifically inhibiting 3' cleavage by the cleavage and polyadenylation specificity factor. Expression of U1A in excess of U1 snRNA causes inhibition of SMN polyadenylation and decreases SMN protein levels. This work reveals a new mechanism for regulating SMN levels and provides new insight into the roles of U1A in 3' processing of mRNAs.
运动神经元存活(SMN)蛋白表达不足会导致脊髓性肌萎缩症,这是一种以运动神经元丧失为特征的神经退行性疾病。尽管维持足够的 SMN 水平很重要,但人们对控制 SMN 表达的因素知之甚少,特别是 SMN 前体 mRNA 的 3' 端加工。在这项研究中,我们确定 U1A 蛋白是 SMN 表达的关键调节剂。U1A 是 U1 snRNP 的一个组成部分,已知其通过直接结合 mRNA 来抑制多聚腺苷酸化。我们表明,U1A 直接结合并具有高亲和力和特异性,与多聚腺苷酸化位点相邻的 SMN 3'-UTR 结合,而不依赖于 U1 snRNP(U1 小核核糖核蛋白)。U1A 的结合通过特异性抑制切割和多聚腺苷酸化特异性因子的 3' 切割来抑制 SMN 前体 mRNA 的多聚腺苷酸化。U1A 的表达超过 U1 snRNA 会导致 SMN 多聚腺苷酸化受到抑制,SMN 蛋白水平降低。这项工作揭示了一种调节 SMN 水平的新机制,并为 U1A 在 mRNA 3' 加工中的作用提供了新的见解。