Structural basis for recognition of ubiquitinated cargo by Tom1-GAT domain

Tom1 (Target of Myb1) is suggested to be involved in the transport of ubiquitinated proteins, through the interaction of its GAT (GGA and Tom1) domain with ubiquitin. Here, we demonstrate that the three-helix bundle of Tom1-GAT has two ubiquitin-binding sites recognizing the hydrophobic Ile44 surface of ubiquitin. The complex crystal structure demonstrates that the first site is a hydrophobic patch on helices alpha 1 and alpha 2. NMR and biochemical data revealed that the N-terminal half of helix alpha 3 of Tom1-GAT constitutes the second, stronger binding site. The double-sided ubiquitin binding enhances the efficiency of recognition of ubiquitinated proteins by Tom1. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.