Cloning and characterization of a cDNA encoding a protein synthesis initiation factor-2 alpha (eIF-2 alpha) kinase from Drosophila melanogaster - Homology to yeast GCN2 protein kinase

Phosphorylation of the alpha subunit of the eukaryotic initiation factor 2 (eIF-2 alpha) is one of the best-characterized mechanisms for downregulating protein synthesis in mammalian cells in response to various stress conditions. In Drosophila, such a regulatory mechanism has not been elucidated, We report the molecular cloning and characterization of DGCN2, a Drosophila eIF-2 alpha kinase related to yeast GCN2 protein kinase, DGCN2 contains all of the 12 catalytic subdomains characteristic of eukaryotic Ser/Thr protein kinases and the conserved sequence of eIF-2 alpha kinases in subdomain V. A large insert of 94 amino acids, which is characteristic of eIF-2 alpha kinases, is also present between subdomains TV and V. It is particularly notable that DGCN2 possesses an amino acid sequence related to class II aminoacyl-tRNA synthetases, a unique feature of yeast GCN2 protein kinase, DGCN2 expression is developmentally regulated, During embryogenesis, DGCN2 mRNA is dynamically expressed in several tissues, Interestingly, at later stages this expression becomes restricted to a few cells of the central nervous system, Affinity-purified antibodies, raised against a synthetic peptide based on the predicted DGCN2 sequence, specifically immunoprecipitated an eIF-2 alpha kinase activity and recognized an similar to 175 kDa phosphoprotein in Western blots of Drosophila embryo extracts.