Cryo-EM Structure of Saccharomyces cerevisiae Target of Rapamycin Complex 2

The Target Of Rapamycin (TOR) kinase assembles into two, distinct multiprotein complexes, conserved across eukaryote evolution. In contrast to TOR Complex1 (TORC1), TORC2 kinase activity is not inhibited by the macrolide rapamycin. Here, we present the structure of Saccharomyces cerevisiae TORC2 determined by electron cryo-microscopy. TORC2 contains six subunits assembling into a 1.4 MDa rhombohedron. Tor2 and Lst8 form the common core of both TOR complexes. Avo3/Rictor is unique to TORC2, but interacts with the same HEAT repeats of Tor2 that are engaged by Kog1/Raptor in mammalian TORC1, explaining the mutual exclusivity of these two proteins. Density, which we conclude is Avo3, occludes the FKBP12-rapamycin-binding site of Tor2’s FRB-domain rendering TORC2 rapamycin-insensitive and recessing the kinase active-site. Although mobile, Avo1/hSin1 further restricts access to the active site as its Conserved-Region-In-the-Middle (CRIM) domain is positioned along an edge of the TORC2 active-site-cleft, consistent with a role for CRIM in substrate recruitment.