Mauron A, Spohr G
Eur J Biochem. 1978 Jan 16;82(2):619-25. doi: 10.1111/j.1432-1033.1978.tb12058.x.
The turn-over of cytoplasmic messenger-like RNA not associated with polyribosomes as well as that of polyribosomal mRNA was investigated by labelling with [3H]uridine in conditions of arrested ribosomal RNA and mitochondrial RNA synthesis. The synthesis of ribosomal RNA was inhibited with toyokamycin and that of mitochondrial RNA with ethidium bromide. In both accumulation kinetics and actinomycin-D-chase experiments, cytoplasmic messenger-like ribonucleoprotein particles and polyribosomes were fractionated by buoyant density centrifugation in CsCl gradients. The half-life of free m1RNA was found to be of 1--2 h whereas the bulk of polyribosomal mRNA was stable over the time period considered (up to 8 h) but with a minor short-lived component. Purification of RNA from polyribosomes labelled under the same conditions and fractionation of it into polyadenylated and non-polyadenylated fractions showed that this short-lived minor component of half-life less than 1 h is non-polyadenylated.
在核糖体RNA和线粒体RNA合成受阻的条件下,用[3H]尿苷标记,研究了与多核糖体无关的细胞质信使样RNA以及多核糖体mRNA的周转情况。用丰加霉素抑制核糖体RNA的合成,用溴化乙锭抑制线粒体RNA的合成。在累积动力学和放线菌素D追踪实验中,通过在CsCl梯度中进行浮力密度离心,对细胞质信使样核糖核蛋白颗粒和多核糖体进行分级分离。发现游离m1RNA的半衰期为1-2小时,而在考虑的时间段内(长达8小时),大部分多核糖体mRNA是稳定的,但有一小部分寿命较短的成分。从在相同条件下标记的多核糖体中纯化RNA,并将其分为多聚腺苷酸化和非多聚腺苷酸化部分,结果表明,这种半衰期小于1小时的寿命较短的小成分是非多聚腺苷酸化的。