RRC ID 53879
Author Fukunaga R, Hunter T.
Title MNK1, a new MAP kinase-activated protein kinase, isolated by a novel expression screening method for identifying protein kinase substrates.
Journal EMBO J
Abstract We have developed a novel expression screening method for identifying protein kinase substrates. In this method, a lambda phage cDNA expression library is screened by in situ, solid-phase phosphorylation using purified protein kinase and [gamma-32P]ATP. Screening a HeLa cDNA library with ERK1 MAP kinase yielded cDNAs of previously characterized ERK substrates, c-Myc and p90RSK, demonstrating the utility of this method for identifying physiological protein kinase substrates. A novel clone isolated in this screen, designated MNK1, encodes a protein-serine/threonine kinase, which is most similar to MAP kinase-activated protein kinase 2 (MAPKAP-K2), 3pK/MAPKAP-K3 and p90RSK. Bacterially expressed MNK1 was phosphorylated and activated in vitro by ERK1 and p38 MAP kinases but not by JNK/SAPK. Further, MNK1 was activated upon stimulation of HeLa cells with 12-O-tetradecanoylphorbol-13-acetate, fetal calf serum, anisomycin, UV irradiation, tumor necrosis factor-alpha, interleukin-1beta, or osmotic shock, and the activation by these stimuli was differentially inhibited by the MEK inhibitor PD098059 or the p38 MAP kinase inhibitor SB202190. Together, these results indicate that MNK1 is a novel class of protein kinase that is activated through both the ERK and p38 MAP kinase signaling pathways.
Volume 16(8)
Pages 1921-33
Published 1997-4-15
DOI 10.1093/emboj/16.8.1921
PMID 9155018
PMC PMC1169795
MeSH Amino Acid Sequence Animals Base Sequence Calcium-Calmodulin-Dependent Protein Kinases / antagonists & inhibitors Calcium-Calmodulin-Dependent Protein Kinases / metabolism* Cell Line Cloning, Molecular / methods* DNA, Complementary / genetics Enzyme Activation Enzyme Inhibitors / pharmacology Flavonoids / pharmacology HeLa Cells Humans Imidazoles / pharmacology Intracellular Signaling Peptides and Proteins Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases* Molecular Sequence Data Phosphorylation Protein-Serine-Threonine Kinases / genetics Protein-Serine-Threonine Kinases / metabolism* Pyridines / pharmacology Recombinant Fusion Proteins Sequence Homology, Amino Acid Signal Transduction / physiology Spodoptera Substrate Specificity p38 Mitogen-Activated Protein Kinases
IF 9.889
Times Cited 518
DNA material Lambda GEX5 (RDB01911)