Cardiovascular and Aging Research, Department of Medicine, Division of Physiology, University of Fribourg, Chemin du Musée 5, 1700, Fribourg, Switzerland.
National Center of Competence in Research "Kidney.CH", Zurich, Switzerland.
Cell Death Dis. 2018 Feb 22;9(3):313. doi: 10.1038/s41419-018-0356-9.
Type-II L-arginine:ureahydrolase, arginase-II (Arg-II), is shown to activate mechanistic target of rapamycin complex 1 (mTORC1) pathway and contributes to cell senescence and apoptosis. In an attempt to elucidate the underlying mechanism, we identified myosin-1b (Myo1b) as a mediator. Overexpression of Arg-II induces re-distribution of lysosome and mTOR but not of tuberous sclerosis complex (TSC) from perinuclear area to cell periphery, dissociation of TSC from lysosome and activation of mTORC1-ribosomal protein S6 kinase 1 (S6K1) pathway. Silencing Myo1b prevents all these alterations induced by Arg-II. By overexpressing Myo1b or its mutant with point mutation in its pleckstrin homology (PH) domain we further demonstrate that this effect of Myo1b is dependent on its PH domain that is required for Myo1b-lysosome association. Notably, Arg-II promotes association of Myo1b with lysosomes. In addition, we show that in senescent vascular smooth muscle cells with elevated endogenous Arg-II, silencing Myo1b prevents Arg-II-mediated lysosomal positioning, dissociation of TSC from lysosome, mTORC1 activation and cell apoptosis. Taken together, our study demonstrates that Myo1b mediates the effect of Arg-II in activating mTORC1-S6K1 through promoting peripheral lysosomal positioning, that results in spatial separation and thus dissociation of TSC from lysosome, leading to hyperactive mTORC1-S6K1 signaling linking to cellular senescence/apoptosis.
II 型 L-精氨酸:尿素水解酶,精氨酸酶-II(Arg-II)被证明可激活雷帕霉素靶蛋白复合物 1(mTORC1)途径,并促进细胞衰老和凋亡。为了阐明潜在的机制,我们鉴定出肌球蛋白-1b(Myo1b)作为一种介质。Arg-II 的过表达诱导溶酶体和 mTOR 的重新分布,但不会使结节性硬化复合物(TSC)从核周区域转移到细胞外周,TSC 从溶酶体解离以及 mTORC1-核糖体蛋白 S6 激酶 1(S6K1)途径的激活。沉默 Myo1b 可防止 Arg-II 诱导的所有这些变化。通过过表达 Myo1b 或其在其 pleckstrin 同源(PH)结构域中有点突变的突变体,我们进一步证明了 Myo1b 的这种作用依赖于其 PH 结构域,该结构域对于 Myo1b-溶酶体的结合是必需的。值得注意的是,Arg-II 促进了 Myo1b 与溶酶体的结合。此外,我们还表明,在衰老的血管平滑肌细胞中,内源性 Arg-II 水平升高,沉默 Myo1b 可防止 Arg-II 介导的溶酶体定位、TSC 从溶酶体解离、mTORC1 激活和细胞凋亡。总之,我们的研究表明,Myo1b 通过促进周围溶酶体定位来介导 Arg-II 激活 mTORC1-S6K1 的作用,从而导致 TSC 从溶酶体解离,mTORC1-S6K1 信号转导过度活跃,与细胞衰老/凋亡相关。