Cryo-EM Structure of Nucleotide-Bound Tel1^ATM Unravels the Molecular Basis of Inhibition and Structural Rationale for Disease-Associated Mutations

Yeast Tel1 and its highly conserved human ortholog ataxia-telangiectasia mutated (ATM) are large protein kinases central to the maintenance of genome integrity. Mutations in ATM are found in ataxia-telangiectasia (A-T) patients and ATM is one of the most frequently mutated genes in many cancers. Using cryoelectron microscopy, we present the structure of Tel1 in a nucleotide-bound state. Our structure reveals molecular details of key residues surrounding the nucleotide binding site and provides a structural and molecular basis for its intrinsically low basal activity. We show that the catalytic residues are in a productive conformation for catalysis, but the phosphatidylinositol 3-kinase-related kinase (PIKK) regulatory domain insert restricts peptide substrate access and the N-lobe is in an open conformation, thus explaining the requirement for Tel1 activation. Structural comparisons with other PIKKs suggest a conserved and common allosteric activation mechanism. Our work also provides a structural rationale for many mutations found in A-T and cancer. Yates et al. describe the cryo-EM structure of Tel1 kinase in a nucleotide-bound state. The 3.7 Å structure provides a structural basis for its intrinsically low basal activity and a potential allosteric activation mechanism, and further provides insight into the occurrence of many disease-associated mutations of its human ortholog ATM.