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首页> 外文期刊>Molecular and Cellular Biology >Molecular cloning and sequence of a cholesterol-repressible enzyme related to prenyltransferase in the isoprene biosynthetic pathway.
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Molecular cloning and sequence of a cholesterol-repressible enzyme related to prenyltransferase in the isoprene biosynthetic pathway.

机译：异戊二烯生物合成途径中与异戊二烯基转移酶相关的胆固醇可抑制酶的分子克隆和序列。

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Differential hybridization and molecular cloning have been used to isolate CR39, a cDNA which hybridizes to a 1.2-kilobase (kb) mRNA in rat liver. The level of CR39 mRNA was increased seven- to ninefold over normal levels by dietary cholestyramine and mevinolin and decreased about fourfold compared with normal levels by cholesterol feeding or administration of mevalonate. Similar changes in the mRNA levels of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and HMG-CoA synthase were observed under the various conditions. In vitro translation of either CR39 hybrid selected RNA or 1.2-kb CR39 RNA generated by an SP6 in vitro transcription system produced a polypeptide of 39,000 daltons. As deduced from the nucleotide sequence of a full-length CR39 cDNA, the rat CR39 polypeptide contained 344 amino acids and had a molecular weight of 39,615. The predicted amino acid composition and submit molecular weight of the rat CR39 were very similar to those of prenyltransferases isolated from chicken, pig, and human. The sequence of amino acid residues 173 through 203 in the rat CR39 polypeptide showed that 17 out of 30 matched an active-site peptide of avian liver prenyltransferase. Thus, alterations in the rate of cholesterogenesis resulted in the coordinate regulation of three mRNAs encoding HMG-CoA reductase, HMG-CoA synthase, and CR39, the latter being tentatively identified as prenyltransferase.
机译：差异杂交和分子克隆已用于分离CR39，它是一种与大鼠肝脏中1.2千碱基（kb）mRNA杂交的cDNA。饮食中的胆甾醇胺和美维宁使CR39 mRNA的水平比正常水平增加了七到九倍，而胆固醇喂养或甲羟戊酸的给药则使CR39 mRNA的水平下降了约四倍。在各种条件下，观察到3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶和HMG-CoA合酶的mRNA水平发生类似变化。由SP6体外转录系统产生的CR39杂种选择RNA或1.2-kb CR39 RNA的体外翻译产生了39,000道尔顿的多肽。从全长CR39 cDNA的核苷酸序列推导，大鼠CR39多肽含有344个氨基酸，分子量为39,615。大鼠CR39的预测氨基酸组成和呈递分子量与从鸡，猪和人中分离的异戊二烯基转移酶非常相似。大鼠CR39多肽中氨基酸残基173至203的序列显示，30个中的17个与禽肝异戊二烯基转移酶的活性位点肽匹配。因此，胆甾醇生成速率的改变导致了三个编码HMG-CoA还原酶，HMG-CoA合酶和CR39的mRNA的协调调节，后者被暂时鉴定为异戊二烯基转移酶。

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