Phosphorylation of eukaryotic initiation factor (eIF) 2 alpha and inhibition of eIF-2B in GH3 pituitary cells by perturbants of early protein processing that induce GRP78.
Agents that mobilize sequestered intracellular Ca2+, including ionophore A23187, EGTA, thapsigargin, and Cbz-Gly-Phe-NH2 (where Cbz is benzyloxycarbonyl), or mild reducing agents, such as dithiothreitol, disrupt early protein processing in the endoplasmic reticulum (ER), inhibit translational initiation, and trigger the induction of GRP78, an ER resident protein. Inhibition of translational initiation in response to acute treatment (15-30 min) of intact GH3 pituitary cells with each of these agents was accompanied by an average 5-fold increase in the amount of phosphorylated eukaryotic initiation factor (eIF) 2 alpha and a 50% reduction in eIF-2B activity. With continued exposure to A23187 (3 h) rates of amino acid incorporation partially recovered, eIF-2 alpha became dephosphorylated, and the inhibition of eIF-2B activity was abolished. These chronic effects were blocked by actinomycin D. Accumulating evidence that the ER may regulate rates of translational initiation through a signaling system altering the activity of eIF-2 is discussed.
Prostko
C R
CR
Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854.
Brostrom
M A
MA
Malara
E M
EM
Brostrom
C O
CO
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D004533:Egtazic Acid / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D018832:Molecular Chaperones; D010766:Phosphorylation; D010911:Pituitary Neoplasms; D014176:Protein Biosynthesis / Q000187:drug effects; D011506:Proteins / Q000037:antagonists & inhibitors*; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin
D002273:Carcinogens; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D015852:Eukaryotic Initiation Factor-2; D020662:Guanine Nucleotide Exchange Factors; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011506:Proteins; D013729:Terpenes; D000001:Calcimycin; D004533:Egtazic Acid; D019284:Thapsigargin