title: BCR-ABL residues interacting with ponatinib are critical to preserve the tumorigenic potential of the oncoprotein.
abstract: Patients with chronic myeloid leukemia in whom tyrosine kinase inhibitors (TKIs) fail often present mutations in the BCR-ABL catalytic domain. We noticed a lack of substitutions involving 4 amino acids (E286, M318, I360, and D381) that form hydrogen bonds with ponatinib. We therefore introduced mutations in each of these residues, either preserving or altering their physicochemical properties. We found that E286, M318, I360, and D381 are dispensable for ABL and BCR-ABL protein stability but are critical for preserving catalytic activity. Indeed, only a "conservative" I360T substitution retained kinase proficiency and transforming potential. Molecular dynamics simulations of BCR-ABL(I360T) revealed differences in both helix αC dynamics and protein-correlated motions, consistent with a modified ATP-binding pocket. Nevertheless, this mutant remained sensitive to ponatinib, imatinib, and dasatinib. These results suggest that changes in the 4 BCR-ABL residues described here would be selected against by a lack of kinase activity or by maintained responsiveness to TKIs. Notably, amino acids equivalent to those identified in BCR-ABL are conserved in 51% of human tyrosine kinases. Hence, these residues may represent an appealing target for the design of pharmacological compounds that would inhibit additional oncogenic tyrosine kinases while avoiding the emergence of resistance due to point mutations.
authors.affiliation: 2P.G.V., Department of Clinical and Molecular Biomedicine, University of Catania, Via Androne, 85, 95124 Catania, Italy. pvigneri@libero.it.
authors.forename: Pietro
authors.initials: P
authors.lastname: Buffa
authors.forename: Chiara
authors.initials: C
authors.lastname: Romano
authors.forename: Alessandro
authors.initials: A
authors.lastname: Pandini
authors.forename: Michele
authors.initials: M
authors.lastname: Massimino
authors.forename: Elena
authors.initials: E
authors.lastname: Tirrò
authors.forename: Francesco
authors.initials: F
authors.lastname: Di Raimondo
authors.forename: Livia
authors.initials: L
authors.lastname: Manzella
authors.forename: Franca
authors.initials: F
authors.lastname: Fraternali
authors.forename: Paolo G
authors.initials: PG
authors.lastname: Vigneri
mesh_terms: D001483:Base Sequence; D055162:Biocatalysis; D002460:Cell Line; D017931:DNA Primers; D016044:Fusion Proteins, bcr-abl / Q000378:metabolism*; D006801:Humans; D007093:Imidazoles / Q000378:metabolism*; D011724:Pyridazines / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D007093:Imidazoles; D011724:Pyridazines; C545373:ponatinib; D016044:Fusion Proteins, bcr-abl
keywords: CML; TKIs; molecular dynamics; mutations; resistance