Lovastatin suppresses erythropoietin receptor surface expression through dual inhibition of glycosylation and geranylgeranylation.
Erythropoietin (Epo) is a cytokine that is required for the survival of erythroid progenitors through interaction with its receptor on the surface of these cells. Recent studies showed that erythropoietin receptor (EpoR) is expressed on many cancer cells. The factors that govern EpoR expression on the cell surface are poorly understood. Using both biotinlyation and radiolabeled Epo binding experiments, we show here that Epo starvation of the Epo-dependent erythroleukemia cell line, ASE2, leads to a time-dependent increase in both forms of EpoR, the maturing 64 kDa and the mature 66 kDa proteins. Mevalonate depletion inhibits the formation of the highly glycosylated mature form of EpoR without affecting the other form. Treatment of cells with lovastatin, a selective inhibitor of the rate-limiting enzyme in the mevalonate pathway leads to inhibition of cell surface EpoR that is induced by Epo starvation. The effect of lovastatin appears to be the consequence of inhibition of two processes, glycosylation and geranylgeranylation. Adding back geranylgeranyl pyrophosphate to lovastatin-treated cells completely prevents the lovastatin effect on EpoR expression. Dolichol, the sugar carrier in N-linked glycosylation that is derived from the mevalonate pathway, partially reverses lovastatin's effect. The glycosylation inhibitor tunicamycin also partially suppresses EpoR surface expression. Inhibiting protein geranylgeranylation mimics the effect of lovastatin and inhibits EpoR surface expression in a concentration-dependent manner. Finally, lovastatin inhibits Epo's stimulatory effects on cell proliferation. These results indicate that mevalonate derivatives are required for normal EpoR expression on the cell surface through two pathways, glycosylation and geranylgeranylation.
Hamadmad
Sumaya N
SN
Department of Pharmacology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, United States.
Hohl
Raymond J
RJ
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000924:Anticholesteremic Agents / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D019921:Biotinylation / Q000379:methods; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004286:Dolichols / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004921:Erythropoietin / Q000494:pharmacology; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007457:Iodine Radioisotopes; D053614:Janus Kinase 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008148:Lovastatin / Q000494:pharmacology*; D008798:Mevalonic Acid / Q000378:metabolism / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011106:Polyisoprenyl Phosphates / Q000378:metabolism / Q000494:pharmacology; D017368:Protein Prenylation / Q000187:drug effects; D017467:Receptors, Erythropoietin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000037:antagonists & inhibitors / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D000924:Anticholesteremic Agents; D000998:Antiviral Agents; D004286:Dolichols; C437105:GGTI 286; D007457:Iodine Radioisotopes; D011106:Polyisoprenyl Phosphates; D017467:Receptors, Erythropoietin; D050799:STAT5 Transcription Factor; D014415:Tunicamycin; D004921:Erythropoietin; D008148:Lovastatin; D053614:Janus Kinase 2; D007930:Leucine; C002963:geranylgeranyl pyrophosphate; D008798:Mevalonic Acid