Alterations in molecular chaperones and eIF2alpha during lung endothelial cell apoptosis.
We have previously demonstrated that inhibition of CAAX carboxyl methylation with AGGC caused redistribution and condensation of the ER molecular chaperones, glucose-regulated protein (GRP)-94 and calnexin; an effect that was attenuated by overexpression of dominant active RhoA. We have also shown that AGGC decreased GRP94 protein level; an effect that was dependent on caspase activity. In the present study, we tested the effects of inhibition of posttranslational processing of CAAX proteins on localization and protein levels of molecular chaperones and phosphorylation and protein level of eIF2alpha. We found that both AGGC, which inhibits CAAX carboxyl methylation, and simvastatin, which inhibits CAAX geranylgeranylation, caused relocalization of GRP94, calnexin, and calreticulin, effects that were not seen during endothelial apoptosis induced by TNF-alpha or ultraviolet (UV) irradiation. These results suggest that posttranslational processing of CAAX proteins is important in maintaining localization of molecular chaperones normally found in the ER. We also noted that AGGC, but not simvastatin, TNF-alpha, or UV irradiation, decreased protein levels of most molecular chaperones. Increased eIF2alpha phosphorylation was observed in the early stages of apoptosis, which was independent of the cause of apoptosis. These results suggest that eIF2alpha phosphorylation is a common early response to apoptosis-inducing stimuli. Interestingly, eIF2alpha protein level was decreased in the late stages of apoptosis induced by AGGC, TNF-alpha, and UV irradiation: an effect that was prevented by caspase inhibition. Thus we speculate that caspase(s)-dependent proteolysis of molecular chaperones and eIF2alpha may be novel signaling pathways of apoptosis. We also speculate that increased eIF2alpha phosphorylation is a defensive response against endothelial cell apoptosis.
Lu
Qing
Q
Department of Medicine, Vascular Research Laboratory, Providence Veterans Affairs Medical Center, Alpert Medical School of Brown University, Providence, Rhode Island 02908, USA.
Jankowich
Matthew
M
Newton
Julie
J
Harrington
Elizabeth O
EO
Rounds
Sharon
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D002417:Cattle; D003545:Cysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D004224:Diterpenes / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D008168:Lung / Q000166:cytology*; D008954:Models, Biological; D018832:Molecular Chaperones / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects / Q000528:radiation effects; D021381:Protein Transport / Q000187:drug effects / Q000528:radiation effects; D019821:Simvastatin / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014466:Ultraviolet Rays
D004224:Diterpenes; D015852:Eukaryotic Initiation Factor-2; D018832:Molecular Chaperones; C095716:N-acetyl-S-geranylgeranyl-cysteine; D014409:Tumor Necrosis Factor-alpha; D019821:Simvastatin; D003545:Cysteine