Roitelman J, Simoni R D
Department of Biological Sciences, Stanford University, California 94305-5020.
J Biol Chem. 1992 Dec 15;267(35):25264-73.
The in vivo turnover rate of the endoplasmic reticulum protein 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in the mevalonate (MVA) pathway, is accelerated when excess MVA or sterols are added to the growth medium of cells. As we have shown recently (Roitelman, J., Bar-Nun, S., Inoue, S., and Simoni, R. D. (1991) J. Biol. Chem. 266, 16085-16091), perturbation of cellular Ca2+ homeostasis abrogates the MVA-accelerated degradation of HMG-CoA reductase and HMGal. Here we show that, in contrast, the sterol-accelerated degradation of HMG-CoA reductase is unaffected by Ca2+ perturbation achieved either by Ca2+ ionophore or by inhibitors of the endoplasmic reticulum Ca(2+)-ATPase. The differential effects of Ca2+ perturbation can be attributed neither to global alteration in protein synthesis nor to inhibition of MVA conversion to sterols. Yet, such manipulations markedly reduce the incorporation of MVA into cellular macromolecules, including prenylated proteins. Furthermore, we directly demonstrate that MVA gives rise to at least two distinct signals, one that is essential to support the effect of sterols and another that operates independently of sterols. Our results indicate that the cellular signals operating in the MVA-accelerated turnover of HMG-CoA reductase are distinct from those involved in the sterol-regulated degradation. A working model for the degradation pathway is proposed.
内质网蛋白3-羟基-3-甲基戊二酰辅酶A(HMG-CoA)还原酶是甲羟戊酸(MVA)途径中的限速酶,当向细胞生长培养基中添加过量的MVA或固醇时,其体内周转率会加快。正如我们最近所表明的(罗伊特尔曼,J.,巴尔-努恩,S.,井上,S.,和西蒙尼,R. D.(1991年)《生物化学杂志》266,16085 - 16091),细胞钙稳态的扰动消除了MVA加速的HMG-CoA还原酶和HMGal的降解。在此我们表明,相比之下,HMG-CoA还原酶的固醇加速降解不受通过钙离子载体或内质网钙(2 +)-ATP酶抑制剂实现的钙扰动的影响。钙扰动的不同效应既不能归因于蛋白质合成的整体改变,也不能归因于MVA向固醇转化的抑制。然而,这种操作显著降低了MVA掺入细胞大分子,包括异戊二烯化蛋白。此外,我们直接证明MVA产生至少两种不同的信号,一种对于支持固醇的作用至关重要,另一种独立于固醇起作用。我们的结果表明,在MVA加速的HMG-CoA还原酶周转中起作用的细胞信号与参与固醇调节降解的信号不同。提出了降解途径的工作模型。