Chan K M, Kurtz D T, Feigelson P
Biochemistry. 1978 Jul 25;17(15):3092-6. doi: 10.1021/bi00608a023.
Alpha2u-globulin is a male rat liver protein under multihormonal control which represents approximately 1% of hepatic protein synthesis. We have measured the rate of transcription of the alpha2u-globulin gene using a nuclear cell-free transcriptional system with mercurated CTP as substrate for the endogenous RNA polymerases. The newly synthesized, mercurated RNA was purified free of endogenous RNA by chromatography on sulfhydryl agarose and hybridized to 3H-labeled alpha2u-globulin cDNA. It was found that, in male rat liver nuclei, alpha2u-globulin RNA sequences represent 0.005% of the total newly synthesized RNA. Actinomycin D or alpha-amanitin completely blocks the synthesis of alpha2u-globulin RNA in these nuclei. No alpha2u-globulin RNA synthesis was detectable in female rat liver nuclei. Thus, within the limits of our detection the absence of hepatic alpha2u-globulin mRNA in female rats appears to be due to a lack of transcription of the alpha2u-globulin gene in these animals.
α2u球蛋白是一种受多种激素调控的雄性大鼠肝脏蛋白,约占肝脏蛋白合成量的1%。我们使用以汞化CTP为内源性RNA聚合酶底物的无细胞细胞核转录系统,测量了α2u球蛋白基因的转录速率。通过在巯基琼脂糖上进行层析,将新合成的汞化RNA与内源性RNA分离并纯化,然后与3H标记的α2u球蛋白cDNA杂交。结果发现,在雄性大鼠肝脏细胞核中,α2u球蛋白RNA序列占新合成RNA总量的0.005%。放线菌素D或α-鹅膏蕈碱可完全阻断这些细胞核中α2u球蛋白RNA的合成。在雌性大鼠肝脏细胞核中未检测到α2u球蛋白RNA的合成。因此,在我们的检测范围内,雌性大鼠肝脏中缺乏α2u球蛋白mRNA似乎是由于这些动物的α2u球蛋白基因缺乏转录。