Regulation of the multiple forms of dopamine beta-hydroxylase by nerve growth factor, dexamethasone, and dibutyryl cyclic AMP in the PC12 pheochromocytoma cell line.
Treatment with nerve growth factor was found to influence the subunit forms of dopamine beta-hydroxylase in PC12 pheochromocytoma cells. In untreated cells, near equal amounts of two subunit forms were observed (apparent Mr = 77,000 and 73,000) by labeling with [35S]methionine. Upon treatment of PC12 cells with nerve growth factor for several days, the Mr = 73,000 subunit form of dopamine beta-hydroxylase was almost exclusively observed. The shift in subunit forms became apparent only after a day of treatment and was maximal with 3 days or more of exposure to nerve growth factor. The dose-response curve was similar to most other nerve growth factor-induced responses in PC12 cells. Neurite outgrowth, however, was not essential for the shift in predominance of the Mr = 73,000 subunit form. This effect of nerve growth factor also occurred in suspension cultures or in the presence of low concentrations of inhibitors of transcription sufficient to prevent neurite outgrowth. Pulse-chase experiments with nerve growth factor-treated cells indicated that the Mr = 77,000 form is initially synthesized (5 min) and is then converted to the Mr = 73,000 form by 30-60 min. Insulin (100 ng/ml) and epidermal growth factor (1 ng/ml) had no effect on the subunit forms of dopamine beta-hydroxylase. However, treatment of PC12 cells for several days with dexamethasone (10(-5)M) or dibutyryl cyclic AMP (1 mM) leads to predominance of the Mr = 73,000 form of the enzyme. These experiments suggest that the proportions of the subunit forms of dopamine beta-hydroxylase can be regulated in cells by external signals and this may reflect alterations in post-translational processing enzymes and may serve as a potential mechanism to regulate catecholamine metabolism.
D000310:Adrenal Gland Neoplasms / Q000201:enzymology*; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology*; D004299:Dopamine beta-Hydroxylase / Q000096:biosynthesis*; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000201:enzymology; D009414:Nerve Growth Factors / Q000494:pharmacology*; D010673:Pheochromocytoma / Q000201:enzymology*; D051381:Rats
D007527:Isoenzymes; D009414:Nerve Growth Factors; D003994:Bucladesine; D003907:Dexamethasone; D004299:Dopamine beta-Hydroxylase