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. Author manuscript; available in PMC: 2016 Jan 22.
Published in final edited form as: Mol Cell. 2014 Dec 18;57(2):207–218. doi: 10.1016/j.molcel.2014.11.013

Figure 2. UVRAG Ser498 is an mTORC1-dependent phosphorylation site.

Figure 2

(A) Alignment of the sequence surrounding UVRAG S498 with those of the known mTORC1 target sites. (B) Rheb induces UVRAG S498 phosphorylation. HA-Rheb was expressed with myc-UVRAG in HEK293T cells. UVRAG S498 phosphorylation was analyzed by WB. (C) UVRAG phosphorylation in MEFs is inhibited by Torin1, Akt inhibitor VIII or HBSS, but not by rapamycin. Immunoprecipitated UVRAG was analyzed for the phosphorylation by WB. (D) The S498 phosphorylation is enhanced by Akt and Rheb. HA-UVRAG was transiently expressed in HEK293T cells alone (−) or together with myc-tagged constitutively active Akt mutant (cAkt) or Rheb. Two days post-transfection, cells were treated with vehicle (−) or Torin1 (+) for 1 h. (E) Deficiency of AMPK, PTEN, or TSC1 in MEFs enhances UVRAG phosphorylation. MEFs were treated with vehicle (−) or Torin1 (+) for 1 h. The UVRAG phosphorylation was analyzed by immunoprecipitation followed by WB. See also Figure S2.