Workman Eileen, Kalda Caitlin, Patel Aalapi, Battle Daniel J
Department of Molecular and Cellular Biochemistry, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
Department of Molecular and Cellular Biochemistry, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210; Department of Neurology, The Ohio State University Wexner Medical Center, Columbus, Ohio 43210.
J Biol Chem. 2015 Jun 19;290(25):15662-15669. doi: 10.1074/jbc.M115.646257. Epub 2015 Apr 24.
Reduced expression of SMN causes spinal muscular atrophy, a severe neurodegenerative disease. Despite the importance of maintaining SMN levels, relatively little is known about the mechanisms by which SMN levels are regulated. We show here that Gemin5, the snRNA-binding protein of the SMN complex, binds directly to the SMN mRNA and regulates SMN expression. Gemin5 binds with high specificity, both in vitro and in vivo, to sequence and structural elements in the SMN mRNA 3'-untranslated region that are reminiscent of the snRNP code to which Gemin5 binds on snRNAs. Reduction of Gemin5 redistributes the SMN mRNA from heavy polysomes to lighter polysomes and monosomes, suggesting that Gemin5 functions as an activator of SMN translation. SMN protein is not stoichiometrically present on the SMN mRNA with Gemin5, but the mRNA-binding activity of Gemin5 is dependent on SMN levels, providing a feedback mechanism for SMN to regulate its own expression via Gemin5. This work both reveals a new autoregulatory pathway governing SMN expression, and identifies a new mechanism through which SMN can modulate specific mRNA expression via Gemin5.
运动神经元存活蛋白(SMN)表达降低会导致脊髓性肌萎缩症,这是一种严重的神经退行性疾病。尽管维持SMN水平很重要,但对于SMN水平受调控的机制却知之甚少。我们在此表明,SMN复合物的小核RNA(snRNA)结合蛋白Gemin5直接与SMN信使核糖核酸(mRNA)结合并调节SMN表达。在体外和体内,Gemin5都以高度特异性结合到SMN mRNA 3'非翻译区的序列和结构元件上,这些元件让人联想到Gemin5在snRNA上结合的小核核糖核蛋白(snRNP)编码。Gemin5的减少会使SMN mRNA从重型多核糖体重新分布到轻型多核糖体和单核糖体,这表明Gemin5作为SMN翻译的激活剂发挥作用。SMN蛋白与Gemin5在SMN mRNA上并非化学计量存在,但Gemin5的mRNA结合活性依赖于SMN水平,这为SMN通过Gemin5调节自身表达提供了一种反馈机制。这项工作既揭示了一种控制SMN表达的新的自我调节途径,又确定了SMN可通过Gemin5调节特定mRNA表达的新机制。