The Rho1 effector Pkc1, but not Bni1, mediates signalling from Tor2 to the actin cytoskeleton

In Saccharomyces cerevisiae, the phosphatidylinositol kinase homologue Tor2 controls the cell-cycle-dependent organisation of the actin cytoskeleton by activating the small GTPase Rho1 via the exchange factor Rom2 [1,2]. Four Rho1 effectors are known, protein kinase C 1 (Pkc1), the formin-family protein Bni1, the glucan synthase Fks and the signalling protein Skn7 [2,3]. Rho1 has been suggested to signal to the actin cytoskeleton via Bni1 and Pkc1; rho1 mutants have never been shown to have defects in actin organisation, however [2,4], We have further investigated the role of Rho1 in controlling actin organisation and have analysed which of the Rho1 effecters mediates Tor2 signalling to the actin cytoskeleton, We show that some, but not all, rho1 temperature-sensitive (rho1(ts)) mutants arrest growth with a disorganised actin cytoskeleton. Both the growth defect and the actin organisation defect of the rho1-2(ts) mutant were suppressed by upregulation of Pkc1 but not by upregulation of Bni1, Fks or Skn7. Overexpression of Pkcl, but not overexpression of Bni1, Fks or Skn7, also rescued a tor2(ts) mutant, and deletion of BNI1 or SKN7 did not prevent the suppression of the tor2(ts) mutation by overexpressed Rom2. Furthermore, overexpression of the Pkc1-controlled mitogen-activated protein (MAP) kinase Mpk1 suppressed the actin defect of tor2(ts) and rho1-2(ts) mutants. Thus, Tor2 signals to the actin cytoskeleton via Rho1, Pkcl and the cell integrity MAP kinase cascade.