Regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase synthesis in Syrian hamster C100 cells by mevinolin, 25-hydroxycholesterol, and mevalonate: The role of posttranscriptional control

Abstract

C100 is a baby hamster kidney cell line that expresses high levels of HMG-CoA reductase relative to its parental cell line SV28. In this study the effects of the oxysterol 25-hydroxycholesterol and mevalonate on the mRNA level and rate of synthesis for HMG-CoA reductase were evaluated in C100 cells treated with mevinolin, a competitive inhibitor of HMG-CoA reductase. The addition of 25-hydroxycholesterol to the cell culture medium resulted in a fourfold decrease in both the rate of synthesis and mRNA level for HMG-CoA reductase. Mevalonate at a concentration of 0.4 mM, when added to mevinolin-treated C100 cells, produced no apparent reduction in HMG-CoA reductase mRNA levels and only a small (25%) decline in HMG-CoA synthesis. Mevalonate (0.4 mM) added to 25-hydroxycholesterol-treated cells resulted in no further reduction in the HMG-CoA reductase mRNA level when compared to cells treated with 25-hydroxycholesterol alone, but produced an additional 30-fold decrease in the rate of HMG-CoA reductase synthesis. Degradation of HMG-CoA reductase was rapid in the presence (t_1/2=1.34 h) or absence (t_1/2=1.17 h) of mevinolin and was not changed significantly by adding either 25-hydroxycholesterol, alone (t_1/2=1.30 h) or both 25-hydroxycholesterol and mevalonate (t_1/2=1.30 h) to mevinolin-treated cells. This study demonstrates that mevalonate and 25-hydroxycholesterol act synergistically in the presence of mevinolin to achieve a greater degree of suppression in the rate of HMG-CoA reductase synthesis than can be accounted for by their individual effects on HMG-CoA reductase mRNA. In addition, the data suggest that mevalonate affects the synthesis of HMG-CoA reductase at a yet unidentified posttranscriptional control site.