PKR-like endoplasmic reticulum kinase (PERK) activation following brain ischemia is independent of unfolded nascent proteins.
Transient global brain ischemia results in an immediate inhibition of protein translation upon reperfusion. During early brain reperfusion protein synthesis is inhibited by alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) phosphorylation by the PKR-like endoplasmic reticulum kinase (PERK). Normally, PERK is held in an inactive, monomeric state by the binding of the endoplasmic reticulum (ER) chaperone GRP78 to the lumenal end of PERK. The prevailing view is that ER stress leads to the accumulation of unfolded proteins in the ER lumen. GRP78 dissociates from PERK to bind these accumulated unfolded proteins, leading to PERK activation, phosphorylation of eIF2alpha, and inhibition of translation. To determine if an increase in unfolded nascent proteins following transient brain ischemia contributes to PERK activation, protein synthesis was blocked by intracerebral injection of anisomycin prior to induction of ischemia. Anisomycin inhibited protein synthesis by over 99% and reduced newly synthesized proteins in the ER to approximately 20% of controls. With an ER nearly devoid of newly synthesized proteins, PERK was still activated and was able to phosphorylate eIF2alpha in CA1 neurons during reperfusion. These data strongly argue that PERK activation is independent of the large increase in unfolded nascent proteins within the ER following transient global brain ischemia.
Department of Emergency Medicine, Wayne State University School of Medicine, Detroit, MI 48201, USA.
T H
TH
Sanderson
M P
MP
Deogracias
K K
KK
Nangia
J
J
Wang
G S
GS
Krause
R
R
Kumar
D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D002545:Brain Ischemia / Q000378:metabolism*; D056547:CA1 Region, Hippocampal / Q000187:drug effects / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D008297:Male; D018832:Molecular Chaperones / Q000096:biosynthesis; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D020318:Rats, Long-Evans; D056811:Unfolded Protein Response; D019892:eIF-2 Kinase / Q000378:metabolism*
D000841:Anisomycin; D015852:Eukaryotic Initiation Factor-2; D018832:Molecular Chaperones; D011500:Protein Synthesis Inhibitors; D019892:eIF-2 Kinase