title: Specific in vitro phosphorylation of plant eIF2alpha by eukaryotic eIF2alpha kinases.
abstract: Phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) is known to be an important translational control mechanism in all eukaryotes with the major exception of plants. Regulation of mammalian and yeast eIF2alpha activity is directly governed by specific phosphorylation on Ser-51. We now demonstrate that recombinant wheat wild-type (51S) but not mutant 51-Ala (51A) protein is phosphorylated by human PKR and yeast GCN2, which are defined eIF2alpha kinases. Further, only wheat wild-type eIF2alpha is a substrate for plant-encoded, double-stranded RNA-dependent kinase (pPKR) activity. Plant PKR and GCN2 phosphorylate recombinant yeast eIF2alpha 51S but not the 51A mutant demonstrating that pPKR has recognition site capability similar to established eIF2alpha kinases. A truncated version of wild-type wheat eIF2alpha containing 51S but not the KGYID motif is not phosphorylated by either hPKR or pPKR suggesting that this putative eIF2alpha kinase docking domain is essential for phosphorylation. Taken together, these results demonstrate the homology among eukaryotic eIF2alpha species and eIF2alpha kinases and support the presence of a plant eIF2alpha phosphorylation pathway.
authors.affiliation: Department of Molecular Biology, University of Wyoming, Laramie 82071-3354, USA.
authors.forename: L Y
authors.initials: LY
authors.lastname: Chang
authors.forename: W Y
authors.initials: WY
authors.lastname: Yang
authors.forename: K
authors.initials: K
authors.lastname: Browning
authors.forename: D
authors.initials: D
authors.lastname: Roth
mesh_terms: D005057:Eukaryotic Cells; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D016296:Mutagenesis; D010766:Phosphorylation; D010940:Plant Proteins / Q000378:metabolism*; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D014908:Triticum; D019892:eIF-2 Kinase / Q000378:metabolism*
chemical_list: D015852:Eukaryotic Initiation Factor-2; D010940:Plant Proteins; D011994:Recombinant Proteins; D012694:Serine; D011494:Protein Kinases; D019892:eIF-2 Kinase