Department of Urology, No. 1 Hospital of China Medical University, Shenyang, Liaoning, People's Republic of China.
Mol Cell Biochem. 2012 Feb;361(1-2):249-57. doi: 10.1007/s11010-011-1110-0. Epub 2011 Nov 5.
Mammalian target of rapamycin (mTOR) controls cell growth and proliferation via the raptor-mTOR (TORC1) and rictor-mTOR (TORC2) protein complexes. The mTORC2 containing mTOR and rictor is thought to be rapamycin insensitive and it is recently shown that both rictor and mTORC2 are essential for the development of both embryonic and extra embryonic tissues. To explore rictor function in the early development of mouse embryos, we disrupted the expression of rictor, a specific component of mTORC2, in mouse fertilized eggs by using rictor shRNA. Our results showed that one-cell stage eggs that were lack of rictor could not enter into the two-cell stage normally. Recent biochemical studies suggests that TORC2 is the elusive PDK2 (3'-phosphoinositide-dependent kinase 2) for AKT/PKB Ser473 phosphorylation, which is deemed necessary for AKT function, so we microinjected AKT-S473A into mouse fertilized eggs to investigate whether AKT-S473A is downstream effector of mTOR.rictor to regulate the mitotic division. Our findings revealed that the rictor induced phosphorylation of AKT in Ser473 is required for TORC2 function in early development of mouse embryos.
哺乳动物雷帕霉素靶蛋白(mTOR)通过 Raptor-mTOR(TORC1)和 Rictor-mTOR(TORC2)蛋白复合物控制细胞生长和增殖。mTORC2 包含 mTOR 和 Rictor,被认为对雷帕霉素不敏感,最近的研究表明,Rictor 和 mTORC2 对于胚胎和胚胎外组织的发育都是必不可少的。为了探索 Rictor 在小鼠胚胎早期发育中的功能,我们使用 Rictor shRNA 破坏了小鼠受精卵中 mTORC2 的特定成分 Rictor 的表达。我们的结果表明,缺乏 Rictor 的一细胞期卵不能正常进入二细胞期。最近的生化研究表明,TORC2 是 AKT/PKB Ser473 磷酸化的难以捉摸的 PDK2(3'-磷酸肌醇依赖性激酶 2),这被认为是 AKT 功能所必需的,因此我们将 AKT-S473A 微注射到小鼠受精卵中,以研究 AKT-S473A 是否是 mTOR.rictor 调节有丝分裂分裂的下游效应物。我们的研究结果表明,Rictor 诱导的 AKT 在 Ser473 上的磷酸化对于 TORC2 在小鼠胚胎早期发育中的功能是必需的。