Inhibition of protein synthesis in cortical neurons during exposure to hydrogen peroxide.
Transient cerebral ischemia, which is accompanied by a sustained release of glutamate and zinc, as well as H(2)O(2) formation during the reperfusion period, strongly depresses protein synthesis. We have previously demonstrated that the glutamate-induced increase in cytosolic Ca(2+) is likely responsible for blockade of the elongation step of protein synthesis, whereas Zn(2+) preferentially inhibits the initiation step. In this study, we provide evidence indicating that H(2)O(2) and thapsigargin mobilized a common intracellular Ca(2+) pool. H(2)O(2) treatment stimulated a slow increase in intracellular Ca(2+), and precluded the effect of thapsigargin on Ca(2+) mobilization. H(2)O(2) stimulated the phosphorylation of both eIF-2alpha and eEF-2, in a time- and dose-dependent manner, suggesting that both the blockade of the elongation and of the initiation step are responsible for the H(2)O(2)-induced inhibition of protein synthesis. However, kinetic data indicated that, at least during the first 15 min of H(2)O(2) treatment, the inhibition of protein synthesis resulted mainly from the phosphorylation of eEF-2. In conclusion, H(2)O(2) inhibits protein translation in cortical neurons by a process that involves the phosphorylation of both eIF-2alpha and eEF-2 and the relative contribution of these two events depends on the duration of H(2)O(2) treatment.
Alirezaei
M
M
Chaire de Neuropharmacologie, INSERM U114, Collège de France, Paris, France.
Marin
P
P
Nairn
A C
AC
Glowinski
J
J
Prémont
J
J
D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007424:Intracellular Fluid / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects; D020652:Peptide Elongation Factor 2 / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011506:Proteins / Q000032:analysis; D019284:Thapsigargin / Q000494:pharmacology; D014966:Xanthenes
D000814:Aniline Compounds; D015852:Eukaryotic Initiation Factor-2; D005456:Fluorescent Dyes; D020652:Peptide Elongation Factor 2; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D014966:Xanthenes; C059715:Fluo-3; D019284:Thapsigargin; D006861:Hydrogen Peroxide; D002118:Calcium