Rab5a activates IRS1 to coordinate IGF-AKT-mTOR signaling and myoblast differentiation during muscle regeneration

Abstract

Rab5 is a master regulator for endosome biogenesis and transport while its in vivo physiological function remains elusive. Here, we find that Rab5a is upregulated in several in vivo and in vitro myogenesis models. By generating myogenic Rab5a-deficient mice, we uncover the essential roles of Rab5a in regulating skeletal muscle regeneration. We further reveal that Rab5a promotes myoblast differentiation and directly interacts with insulin receptor substrate 1 (IRS1), an essential scaffold protein for propagating IGF signaling. Rab5a interacts with IRS1 in a GTP-dependent manner and this interaction is enhanced upon IGF-1 activation and myogenic differentiation. We subsequently identify that the arginine 207 and 222 of IRS1 and tyrosine 82, 89, and 90 of Rab5a are the critical amino acid residues for mediating the association. Mechanistically, Rab5a modulates IRS1 activation by coordinating the association between IRS1 and the IGF receptor (IGFR) and regulating the intracellular membrane targeting of IRS1. Both myogenesis-induced and IGF-evoked AKT-mTOR signaling are dependent on Rab5a. Myogenic deletion of Rab5a also reduces the activation of AKT-mTOR signaling during skeletal muscle regeneration. Taken together, our study uncovers the physiological function of Rab5a in regulating muscle regeneration and delineates the novel role of Rab5a as a critical switch controlling AKT-mTOR signaling by activating IRS1.

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Fig. 1: Specific up-regulation of Rab5a during myoblasts differentiation and in vivo myogenesis.
Fig. 2: Myogenic ablation of Rab5a impairs muscle regeneration.
Fig. 3: Rab5a positively regulates myoblast differentiation.
Fig. 4: Rab5a directly interacts with IRS1.
Fig. 5: Rab5a coordinates the interaction between IGFR and IRS1 during myoblast differentiation.
Fig. 6: Rab5a targeted IRS1 to the intracellular membrane.
Fig. 7: Rab5a promotes AKT-mTOR signaling during myoblast differentiation and muscle regeneration.

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Acknowledgements

We thank Ms. Yanwei Li, Junli Xuan and Guifeng Xiao from the core facility platform of Zhejiang university school of medicine for their technical support. This work was supported by the National Key R&D Program of China (2018YFA0800403), the National Natural Science Foundation of China (31571402, 31671417, 31671479, 31371476), the International Science & Technology Cooperation Program of China (the Ministry of Science and Technology of PRC, 2015DFG32130), grants from Science and Technology Commission of Zhejiang Province (LZ20H060001), China Postdoctoral Science Foundation (2019M662034).

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Cong, X.X., Gao, X.K., Rao, X.S. et al. Rab5a activates IRS1 to coordinate IGF-AKT-mTOR signaling and myoblast differentiation during muscle regeneration. Cell Death Differ 27, 2344–2362 (2020). https://doi.org/10.1038/s41418-020-0508-1

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