Department of Bioscience, National Cardiovascular Center Research Institute, Suita, Osaka, Japan.
Genes Cells. 2010 Nov;15(11):1158-67. doi: 10.1111/j.1365-2443.2010.01446.x.
The formation of muscle fibers involves sequential expression of many proteins that regulate key steps during myoblast-to-myotube transition. Myogenin is a major player in the initiation and maintenance of myogenic differentiation in a mouse myoblast cell line, C2C12. RNA-binding proteins bind to specific target RNA sequences and regulate gene expression in a post-transcriptional manner. This study demonstrates that RNA-binding motif protein 24 (Rbm24) interacts with the 3'-untranslated region of myogenin mRNA and affects its half-life in C2C12 myogenesis. Knockdown of Rbm24 expression by RNA interference significantly decreased myogenin expression associated with the inhibition of myogenesis. In contrast, the overexpression of Rbm24 by stable transfection of a plasmid increased myogenin expression and had a positive effect on myogenic differentiation. Ectopic expression of myogenin was also able to restore myogenic differentiation in Rbm24-knockdown cells. Together, our results suggest that Rbm24 binds to myogenin mRNA and regulates its stability in C2C12 cells. Rbm24 plays a crucial role in myogenic differentiation at least in part through a myogenin-dependent post-transcriptional regulatory pathway.
肌纤维的形成涉及许多蛋白质的顺序表达,这些蛋白质调节成肌细胞向肌管过渡过程中的关键步骤。在小鼠成肌细胞系 C2C12 中,肌球蛋白重链是肌发生起始和维持的主要调控因子。RNA 结合蛋白结合到特定的靶 RNA 序列上,并以转录后方式调节基因表达。本研究表明,RNA 结合基序蛋白 24(Rbm24)与肌球蛋白重链 mRNA 的 3'-非翻译区相互作用,并影响其在 C2C12 肌发生中的半衰期。通过 RNA 干扰敲低 Rbm24 的表达会显著降低与肌生成抑制相关的肌球蛋白重链表达。相比之下,通过稳定转染质粒过表达 Rbm24 会增加肌球蛋白重链的表达,并对肌生成分化产生积极影响。肌球蛋白重链的异位表达也能够恢复 Rbm24 敲低细胞的肌生成分化。总之,我们的结果表明,Rbm24 与肌球蛋白重链 mRNA 结合,并在 C2C12 细胞中调节其稳定性。Rbm24 至少部分通过肌球蛋白重链依赖的转录后调控途径在肌生成分化中发挥关键作用。