Geoghegan T E, Knoop S J, Chitpatima C
Biochem Biophys Res Commun. 1983 Jun 29;113(3):923-33. doi: 10.1016/0006-291x(83)91087-2.
A few mRNAs in S-180 ascites cells are found as non-polysomal messenger ribonucleoprotein particles, whose translation is restricted both in vivo and in vitro. To determine if these mRNPs remain untranslated throughout the cell cycle we obtained S-180 ascites cells enriched in cell cycle stages by centrifugation through ficoll gradients. The "inactive" mRNA species were clearly present in the pre-polysomal fraction from S and pre-S phase cells, however they were not detected in the mRNP fraction from post S phase cells. Hybridization of a cloned cDNA probe for one of these species (P-40) to polysomal RNA from pre- or post-S phase cells demonstrated it was present in substantially higher concentrations in polysomes from post-S phase cells, consistent with its translation after S phase. We also observed that a poly (A)-minus form of the mRNA coding for actin was restricted to post S phase cells.
在S-180腹水细胞中,一些mRNA以非多聚体信使核糖核蛋白颗粒的形式存在,其翻译在体内和体外均受到限制。为了确定这些mRNA颗粒在整个细胞周期中是否都未被翻译,我们通过菲可梯度离心获得了富集于细胞周期各阶段的S-180腹水细胞。“无活性”的mRNA种类在S期和S期前细胞的多聚体前级分中明显存在,然而在S期后细胞的mRNA颗粒级分中未检测到。用针对其中一种mRNA(P-40)的克隆cDNA探针与S期前或S期后细胞的多聚体RNA杂交,结果表明它在S期后细胞的多聚体中的浓度要高得多,这与其在S期后进行翻译相一致。我们还观察到,编码肌动蛋白的mRNA的聚腺苷酸缺失形式仅限于S期后细胞。