Rat liver DT-diaphorase: regulation of functional mRNA levels by 3-methylcholanthrene, trans-stilbene oxide, and phenobarbital.
Rat liver DT-diaphorase: regulation of functional mRNA levels by 3-methylcholanthrene, trans-stilbene oxide, and phenobarbital.
复制标题
大鼠肝脏 DT-黄酶：3-甲基胆蒽、反式二苯乙烯氧化物和苯巴比妥对功能性 mRNA 水平的调节。
DOI：
10.1016/0003-9861(84)90556-3
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发表时间：
1984
影响因子：
3.9
通讯作者：
Pickett,CB
中科院分区：
文献类型：
--
作者：
Williams,JB;Wang,R;Lu,AY;Pickett,CB
从未处理、3-甲基胆蒽-、反式二苯乙烯-和苯巴比妥处理的大鼠中分离的总肝脏poly(A+)- rna在兔网红细胞溶解液系统中被翻译，以确定这些外源物对翻译活性DT-diaphorase mRNA水平的影响。它们的体外翻译系统用兔抗纯化的DT-diaphorase培养的IgG免疫沉淀，并进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳。通过使用金黄色葡萄球菌v -8蛋白酶进行有限的肽定位，证实了放射性标记的免疫沉淀产物为DT-diaphorase。这些定量结果表明，在单次给药后8小时，3-甲基胆蒽导致功能性DT-diaphorase mRNA升高8倍；然而，反式二苯乙烯氧化物和苯巴比妥只产生适度的升高，2至3倍，在功能性的DT-diaphorase mRNA水平。这些数据表明，先前注意到的3-甲基胆蒽给药后DT-diaphorase水平的增加[B]。Höjeberg， K. Blomberg， S. Stenberg和C. Lind (1981)Arch。物化学。生物物理学报，207,205 - 216]可以完全由该蛋白特异性mRNA水平的升高来解释。
Total liver poly(A+)-RNA isolated from untreated, and 3-methylcholanthrene-,trans-stilbene oxide-, and phenobarbital-treated rats has been translated in the rabbit reticulocyte lysate system in order to determine the effect of these xenobiotics on the level of translationally active DT-diaphorase mRNA. Thein vitrotranslation systems were subjected to immunoprecipitation with rabbit IgG raised against purified DT-diaphorase and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identity of the radiolabeled, immunoprecipitated product as DT-diaphorase was confirmed by limited peptide mapping usingStaphylococcus aureusV-8 protease. These quantitation results demonstrate that 3-methylcholanthrene leads to an eightfold elevation in functional DT-diaphorase mRNA at 8 h after a single administration of 3-methylcholanthrene; whereas,trans-stilbene oxide and phenobarbital produced only a modest elevation, two- to three-fold, in the functional DT-diaphorase mRNA level. These data indicate that the increase in the level of DT-diaphorase after 3-methylcholanthrene administration noted previously [B. Höjeberg, K. Blomberg, S. Stenberg, and C. Lind (1981)Arch. Biochem. Biophys.207, 205–216] can be totally accounted for by an elevation in the mRNA level specific for this protein.