Shinji Sayaka, Umezawa Koji, Nihashi Yuma, Nakamura Shunichi, Shimosato Takeshi, Takaya Tomohide
Department of Agriculture, Graduate School of Science and Technology, Shinshu University, Nagano, Japan.
Department of Agricultural and Life Science, Faculty of Agriculture, Shinshu University, Nagano, Japan.
Front Cell Dev Biol. 2021 Jan 11;8:616706. doi: 10.3389/fcell.2020.616706. eCollection 2020.
Herein we report that the 18-base telomeric oligodeoxynucleotides (ODNs) designed from the GG genome promote differentiation of skeletal muscle myoblasts which are myogenic precursor cells. We termed these myogenetic ODNs (myoDNs). The activity of one of the myoDNs, iSN04, was independent of Toll-like receptors, but dependent on its conformational state. Molecular simulation and iSN04 mutants revealed stacking of the 13-15th guanines as a core structure for iSN04. The alkaloid berberine bound to the guanine stack and enhanced iSN04 activity, probably by stabilizing and optimizing iSN04 conformation. We further identified nucleolin as an iSN04-binding protein. Results showed that iSN04 antagonizes nucleolin, increases the levels of p53 protein translationally suppressed by nucleolin, and eventually induces myotube formation by modulating the expression of genes involved in myogenic differentiation and cell cycle arrest. This study shows that bacterial-derived myoDNs serve as aptamers and are potential nucleic acid drugs directly targeting myoblasts.
在此我们报告，从GG基因组设计的18碱基端粒寡脱氧核苷酸（ODN）可促进作为成肌前体细胞的骨骼肌成肌细胞分化。我们将这些成肌ODN称为肌源ODN（myoDN）。其中一种myoDN，即iSN04的活性不依赖于Toll样受体，而是依赖于其构象状态。分子模拟和iSN04突变体显示，第13 - 15位鸟嘌呤的堆积是iSN04的核心结构。生物碱小檗碱与鸟嘌呤堆积结合并增强iSN04活性，可能是通过稳定和优化iSN04构象来实现的。我们进一步鉴定出核仁素是一种iSN04结合蛋白。结果表明，iSN04拮抗核仁素，增加被核仁素翻译抑制的p53蛋白水平，并最终通过调节参与成肌分化和细胞周期停滞的基因表达来诱导肌管形成。这项研究表明，细菌衍生的myoDN作为适体，是直接靶向成肌细胞的潜在核酸药物。
