title: Ancestral resurrection reveals evolutionary mechanisms of kinase plasticity.
abstract: Protein kinases have evolved diverse specificities to enable cellular information processing. To gain insight into the mechanisms underlying kinase diversification, we studied the CMGC protein kinases using ancestral reconstruction. Within this group, the cyclin dependent kinases (CDKs) and mitogen activated protein kinases (MAPKs) require proline at the +1 position of their substrates, while Ime2 prefers arginine. The resurrected common ancestor of CDKs, MAPKs, and Ime2 could phosphorylate substrates with +1 proline or arginine, with preference for proline. This specificity changed to a strong preference for +1 arginine in the lineage leading to Ime2 via an intermediate with equal specificity for proline and arginine. Mutant analysis revealed that a variable residue within the kinase catalytic cleft, DFGx, modulates +1 specificity. Expansion of Ime2 kinase specificity by mutation of this residue did not cause dominant deleterious effects in vivo. Tolerance of cells to new specificities likely enabled the evolutionary divergence of kinases.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.
authors.forename: Conor J
authors.initials: CJ
authors.lastname: Howard
authors.affiliation: Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, United States.
authors.forename: Victor
authors.initials: V
authors.lastname: Hanson-Smith
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.
authors.forename: Kristopher J
authors.initials: KJ
authors.lastname: Kennedy
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, United States.
authors.forename: Chad J
authors.initials: CJ
authors.lastname: Miller
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, United States.
authors.forename: Hua Jane
authors.initials: HJ
authors.lastname: Lou
authors.affiliation: Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, United States.
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Johnson
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, United States.
authors.forename: Benjamin E
authors.initials: BE
authors.lastname: Turk
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.
authors.forename: Liam J
authors.identifier: http://orcid.org/0000-0002-4002-0861
authors.initials: LJ
authors.lastname: Holt
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism; D019143:Evolution, Molecular; D006801:Humans; D007700:Kinetics; D008540:Meiosis; D008969:Molecular Sequence Data; D009154:Mutation; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D010802:Phylogeny; D011392:Proline / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D016213:Cyclins; D010455:Peptides; D001120:Arginine; D011392:Proline; D011494:Protein Kinases
keywords: Ime2; S. cerevisiae; ancestral reconstruction; biochemistry; cyclin-dependent kinase; evolution; evolutionary biology; genomics; human; mouse; neurospora; phosphoregulatory networks; protein kinase