Brain ischemia and reperfusion activates the eukaryotic initiation factor 2 alpha kinase, PERK

Reperfusion after global brain ischemia results initially in a widespread s uppression of protein synthesis in neurons, which persists in vulnerable ne urons, that is caused by the inhibition of translation initiation as a resu lt of the phosphorylation of the a-subunit of eukaryotic initiation factor 2 (elF2 alpha). To identify kinases responsible for elF2 alpha phosphorylat ion [elF2 alpha (P)] during brain reperfusion, we induced ischemia by bilat eral carotid artery occlusion followed by post-ischemic assessment of brain elF2 alpha (P) in mice with homozygous functional knockouts in the genes e ncoding the heme-regulated elF2 alpha kinase (HRI), or the amino acid-regul ated elF2 alpha kinase (GCN2). A 10-fold increase in elF2 alpha (P) was obs erved in reperfused wild-type mice and in the HRI-/- or GCN2-/- mice. Howev er, in all reperfused groups, the RNA-dependent protein kinase (PKR)-like e ndoplasmic reticulum elF2 alpha kinase (PERK) exhibited an isoform mobility shift on SDS-PAGE, consistent with the activation of the kinase, These dat a indicate that neither HRI nor GCN2 are required for the large increase in post-ischemic brain elF2 alpha (P), and in conjunction with our previous r eport that elF2 alpha (P) is produced in the brain of reperfused PKR-/- mic e, provides evidence that PERK is the kinase responsible for elF2 alpha. ph osphorylation in the early post-ischemic brain.