Queens College of the City University of New York, Biology Department, 65-30 Kissena Boulevard, Flushing, NY 11367, USA.
Dev Biol. 2012 Dec 1;372(1):103-10. doi: 10.1016/j.ydbio.2012.08.013. Epub 2012 Aug 28.
The Exon Junction Complex (EJC) plays a critical role in multiple posttranscriptional events, including RNA subcellular localization, nonsense-mediated decay (NMD), and translation. We previously reported that knockdown of the EJC core component Eukaryotic initiation factor 4a3 (Eif4a3) results in full-body paralysis of embryos of the frog, Xenopus laevis. Here, we explore the cellular and molecular mechanisms underlying this phenotype. We find that cultured muscle cells derived from Eif4a3 morphants do not contract, and fail to undergo calcium-dependent calcium release in response to electrical stimulation or treatment with caffeine. We show that ryr (ryanodine receptor) transcripts are incorrectly spliced in Eif4a3 morphants, and demonstrate that inhibition of Xenopus Ryr function similarly results in embryonic paralysis. These results suggest that the EJC mediates muscle cell function via regulation of pre-mRNA splicing during early vertebrate embryogenesis.
外显子连接复合物 (EJC) 在多种转录后事件中发挥着关键作用,包括 RNA 的亚细胞定位、无意义介导的衰变 (NMD) 和翻译。我们之前的研究报告表明,EJC 核心成分真核起始因子 4a3 (Eif4a3) 的敲低会导致青蛙非洲爪蟾的胚胎全身瘫痪。在这里,我们探索了这种表型的细胞和分子机制。我们发现,源自 Eif4a3 突变体的培养肌肉细胞不会收缩,并且在电刺激或咖啡因处理时不能响应钙离子依赖性钙离子释放。我们表明,在 Eif4a3 突变体中 ryr(ryanodine receptor)转录本发生了错误剪接,并证明抑制非洲爪蟾 Ryr 功能也会导致胚胎瘫痪。这些结果表明,EJC 通过调节早期脊椎动物胚胎发生过程中的前体 mRNA 剪接来介导肌肉细胞功能。