The Saccharomyces cerevisiae mevalonate diphosphate decarboxylase is essential for viability, and a single Leu-to-Pro mutation in a conserved sequence leads to thermosensitivity
The Saccharomyces cerevisiae mevalonate diphosphate decarboxylase is essential for viability, and a single Leu-to-Pro mutation in a conserved sequence leads to thermosensitivity
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DOI：
10.1128/jb.179.15.4664-4670.1997
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发表时间：
1997-08-01
影响因子：
3.2
通讯作者：
Karst, F
中科院分区：
文献类型：
--
作者：
Berges, T;Guyonnet, D;Karst, F
甲羟戊酸二磷酸脱羧酶是将甲羟戊酸二磷酸转化为异戊烯基二磷酸（类异戊二烯的结构单元）的酶。我们使用了先前描述的甲羟戊酸二磷酸脱羧酶缺陷的酿酒酵母温度敏感突变体（C。Chambon，V. Ladeveve，M.塞尔武斯湖布兰查德角Javelot，B. Vladescu和F.第二十六章：你是谁633-636，1991）来表征突变的等位基因，我们表明保守氨基酸中的单一变化解释了突变体的温度敏感性表型。在erg 19突变的背景和其中ERG 19基因（其被证明是酵母的必需基因）被破坏的菌株中进行了互补实验。野生型甲羟戊酸二磷酸脱羧酶的表位标记允许我们通过通用的一步免疫沉淀程序分离活性形式的酶。此外，在本研究过程中，我们观察到野生型ERG 19基因的高水平表达导致与野生型菌株相比较低的甾醇稳态积累，提示该酶可能是甲羟戊酸途径调控中的关键酶。
The mevalonate diphosphate decarboxylase is an enzyme which converts mevalonate diphosphate to isopentenyl diphosphate, the building block of isoprenoids. We used the Saccharomyces cerevisiae temperature-sensitive mutant defective for mevalonate diphosphate decarboxylase previously described (C. Chambon, V. Ladeveve, M. Servouse, L. Blanchard, C. Javelot, B. Vladescu, and F. Karst, Lipids 26:633-636, 1991) to characterize the mutated allele, We showed that a single change in a conserved amino acid accounts for the temperature-sensitive phenotype of the mutant, Complementation experiments were done both in the erg19-mutated background and in a strain in which the ERG19 gene, which was shown to be an essential gene for yeast, was disrupted, Epitope tagging of the wild-type mevalonate diphosphate decarboxylase allowed us to isolate the enzyme in an active form by a versatile one-step immunoprecipitation procedure, Furthermore, during the course of this study, we observed that a high level of expression of the wild-type ERG19 gene led to a lower sterol steady-state accumulation compared to that of a wild-type strain, suggesting that this enzyme may be a key enzyme in mevalonate pathway regulation.