Rab-Family GTPase Regulates TOR Complex 2 Signaling in Fission Yeast

Background From yeast to human TOR (target of rapamycin) kinase plays pivotal roles in coupling extracellular stimuli to cell growth and metabolism TOR kinase functions in two distinct protein complexes, TOR complex 1 (TORC1) and 2 (TORC2), which phosphorylate and activate different AGC-family protein kinases TORC1 is controlled by the small GTPase Rheb, but little is known about TORC2 regulators Results We have identified the Ryh1 GTPase, a human Rab6 ortholog, as an activator of TORC2 signaling in the fission yeast Schizosaccharomyces pombe Mutational inactivation of Ryh1 or its guanine nucleotide exchange factor compromises the TORC2-dependent phosphorylation of the AGC-family Gad8 kinase In addition, the effector domain of Ryh1 is important for its physical interaction with TORC2 and for stimulation of TORC2 signaling Thus, GTP-bound Ryh1 is likely to be the active form stimulatory to TORC2-Gad8 signaling Consistently, expression of the GTP-locked mutant Ryh1 is sufficient to promote interaction between TORC2 and Gad8 and to induce Gad8 hyperphosphorylation The loss of functional Ryh1, TORC2, or Gad8 brings about similar vacuolar fragmentation and stress sensitivity, further corroborating their involvement in a common cellular process Human Rab6 can substitute Ryh1 in S pombe and therefore Rab6 may be a potential activator of TORC2 in mammals Conclusions In its GTP-bound form, Ryh1, an evolutionarily conserved Rab GTPase activates TORC2 signaling to the AGC kinase Gad8 The Ryh1 GTPase and the TORC2-Gad8 pathway are required for vacuolar integrity and cellular stress resistance in S pombe