Differential effect of Mn2+ on the hemin-controlled translational repressor and the double-stranded RNA-activated inhibitor.
The inhibition of protein synthesis that occurs when rabbit reticulocyte lysate is incubated in the absence of hemin is due to the activation of a protein kinase termed the hemin-controlled translational repressor, and that occurring when reticulocyte lysate is incubated with a low level of double-stranded RNA is mediated by the activation of a separate protein kinase termed the double-stranded RNA-activated inhibitor. Both the hemin-controlled translational repressor and the double-stranded RNA-activated inhibitor act by phosphorylating the Mr = 35,000 (alpha) subunit of eIF-2. MnCl2 (0.5 mM) partly reverses the inhibition of protein synthesis produced by hemin deficiency but not that induced by double-stranded RNA. In addition, Mn2+ reverses the inhibition of binding of [35S]Met-tRNAf to reticulocyte ribosomal components, isolated on Sepharose 6B, produced by the hemin-controlled translational repressor but not by the double-stranded RNA-activated inhibitor. The effect of Mn2+ is mediated at the level of activation and eIF-2 alpha kinase activity of these two regulatory protein kinases. Specifically, Mn2+ inhibits activation of the hemin-controlled translational repressor in the absence of hemin and the phosphorylation of eIF-2 alpha by pre-activated translational repressor. In contrast, the phosphorylation of eIF-2 alpha by the double-stranded RNA-activated inhibitor is not suppressed by Mn2+, and the activation and autophosphorylation of this inhibitor is enhanced by Mn2+. Finally, while the activation and inactivation of the hemin-controlled translational repressor does not appear to be mediated by autophosphorylation and dephosphorylation, the activation of the double-stranded RNA-activated inhibitor does appear to require autophosphorylation.
D000818:Animals; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2; D006418:Heme / Q000494:pharmacology*; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology*; D010448:Peptide Initiation Factors / Q000378:metabolism; D010766:Phosphorylation; D014176:Protein Biosynthesis / Q000187:drug effects*; D011494:Protein Kinases / Q000097:blood*; D011506:Proteins / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000235:genetics*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism*
D015852:Eukaryotic Initiation Factor-2; D010448:Peptide Initiation Factors; D011506:Proteins; D012330:RNA, Double-Stranded; D006418:Heme; D008345:Manganese; D011494:Protein Kinases