Wheat germ initiation factor 2 (WG x eIF2) decreases the inhibition in protein synthesis and eIF2B activity of reticulocyte lysates mediated by eIF2alpha phosphorylation.

Phosphorylation of serine 51 residue in the alpha-subunit of eukaryotic initiation factor 2 (eIF2alpha) impairs the guanine nucleotide exchange (GNE) activity of eIF2B protein and thereby inhibits protein synthesis in mammalian systems, insects and yeast. It is not known if phosphorylation of plant eIF2 can inhibit an eIF2B-like activity. Interestingly purified wheat germ eIF2 (WG x eIF2) can exchange guanine nucleotides in vitro without the addition of any protein factor like eIF2B. It is not clear if this is due to a contaminant eIF2B-like activity associated with WG x eIF2 or because the affinity of WG x eIF2 for GDP and GTP is not markedly different. Our observations here indicate that the GNE activity of WG x eIF2 is not inhibited upon phosphorylation of the p41-42 doublet subunit in WG x eIF2 by reticulocyte eIF2alpha kinases, or in the presence of reticulocyte eIF2(alphaP) in which serine 51 residue is phosphorylated. Further, addition of WG x eIF2 reduces the inhibition in eIF2B activity, protein synthesis, and also the formation of 15S complex that occurs between reticulocyte eIF2(alphaP) and eIF2B protein in heme-deficient or poly(IC)-treated reticulocyte lysates, presumably by a mechanism of competition between wheat germ and reticulocyte eIF2 for phosphorylation. Unlike reticulocyte eIF2(alphaP), phosphorylated WG x eIF2 is unable to interact with reticulocyte eIF2B to form a 15S complex. The ability of WG x eIF2 to exchange guanine nucleotides independent of an eIF2B like protein and the inability of phosphorylated WG x eIF2 to interact with reticulocyte eIF2B suggests that WG x eIF2 is different from mammalian eIF2 and these differences may have occurred in evolution probably due to some changes in the amino acid sequences around the phosphorylation site in eIF2alpha.