Pickett C B, Lu A Y
Proc Natl Acad Sci U S A. 1981 Feb;78(2):893-7. doi: 10.1073/pnas.78.2.893.
Liver poly(A)+RNA isolated from untreated and phenobarbital-treated rats has been translated in the rabbit reticulocyte cell-fre system in order to determine the level of translationally active epoxide hydrolase (EC 3.3.2.3) mRNA. The in vitro translation systems were immunoprecipitated with rabbit IgG prepared against purified epoxide hydrolase, and the amount of epoxide hydrolase synthesized by the lysate programmed with control and phenobarbital poly(A)+RNA was quantitated. The level of translatable epoxide hydrolase mRNA is increased 3-fold after chronic phenobarbital administration. This level of induction correlates well with the 5-fold induction in catalytic activity of epoxide hydrolase (using styrene 7,8-oxide as substrate) in microsomes isolated from phenobarbital-treated rats. Therefore, we suggest that chronic phenobarbital administration increases the amount of functional epoxide hydrolase in rat liver microsomes by way of an increase in the translatable mRNA level encoding for the enzyme. We do not know whether the increase in mRNA is the result of increased transcription or messenger stability.
从未经处理和经苯巴比妥处理的大鼠中分离出肝脏多聚腺苷酸加尾RNA（poly(A)+RNA），并在兔网织红细胞无细胞系统中进行翻译，以确定具有翻译活性的环氧化物水解酶（EC 3.3.2.3）mRNA的水平。用针对纯化的环氧化物水解酶制备的兔免疫球蛋白G对体外翻译系统进行免疫沉淀，并对用对照和苯巴比妥多聚腺苷酸加尾RNA编程的裂解物合成的环氧化物水解酶量进行定量。长期给予苯巴比妥后，可翻译的环氧化物水解酶mRNA水平增加了3倍。这种诱导水平与从经苯巴比妥处理的大鼠中分离出的微粒体中环氧化物水解酶（以苯乙烯7,8-氧化物为底物）催化活性5倍的诱导密切相关。因此，我们认为长期给予苯巴比妥通过增加编码该酶的可翻译mRNA水平，从而增加大鼠肝脏微粒体中功能性环氧化物水解酶的量。我们不知道mRNA的增加是转录增加还是信使稳定性增加的结果。
