Hampton R Y, Rine J
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
J Cell Biol. 1994 Apr;125(2):299-312. doi: 10.1083/jcb.125.2.299.
Numerous integral membrane proteins are degraded in the mammalian ER. HMG-CoA reductase (HMG-R), a key enzyme in the mevalonate pathway by which isoprenoids and sterols are synthesized, is one substrate of ER degradation. The degradation of HMG-R is modulated by feedback signals from the mevalonate pathway. We investigated the role of regulated degradation of the two isozymes of HMG-R, Hmg1p and Hmg2p, in the physiology of Saccharomyces cerevisiae. Hmg1p was quite stable, whereas Hmg2p was rapidly degraded. Degradation of Hmg2p proceeded independently of vacuolar proteases or secretory traffic, indicating that Hmg2p degradation occurred at the ER. Hmg2p stability was strongly affected by modulation of the mevalonate pathway through pharmacological or genetic means. Decreased mevalonate pathway flux resulted in decreased degradation of Hmg2p. One signal for degradation of Hmg2p was a nonsterol, mevalonate-derived molecule produced before the synthesis of squalene. Genetic evidence indicated that a farnesylated protein may also be necessary for Hmg2p degradation. Studies with reporter genes demonstrated that the stability of each isozyme was determined by its noncatalytic NH2-terminal domain. Our data show that ER protein degradation is widely conserved among eukaryotes, and that feedback control of HMG-R degradation is an ancient paradigm of regulation.
许多整合膜蛋白在哺乳动物内质网中被降解。3-羟基-3-甲基戊二酰辅酶A还原酶(HMG-R)是甲羟戊酸途径中的关键酶,类异戊二烯和甾醇通过该途径合成,它是内质网降解的一个底物。HMG-R的降解受甲羟戊酸途径反馈信号的调节。我们研究了HMG-R的两种同工酶Hmg1p和Hmg2p的调控降解在酿酒酵母生理学中的作用。Hmg1p相当稳定,而Hmg2p迅速降解。Hmg2p的降解独立于液泡蛋白酶或分泌运输进行,表明Hmg2p的降解发生在内质网。通过药理学或遗传学手段调节甲羟戊酸途径,会强烈影响Hmg2p的稳定性。甲羟戊酸途径通量降低导致Hmg2p降解减少。Hmg2p降解的一个信号是在角鲨烯合成之前产生的一种非甾醇、由甲羟戊酸衍生的分子。遗传学证据表明,一种法尼基化蛋白可能也是Hmg2p降解所必需的。对报告基因的研究表明,每种同工酶的稳定性由其非催化性的氨基末端结构域决定。我们的数据表明,内质网蛋白降解在真核生物中广泛保守,并且HMG-R降解的反馈控制是一种古老的调控模式。