Dahnke H G
Hoppe Seylers Z Physiol Chem. 1975 Oct;356(10):1555-63.
The radioactivity of RNA, DNA and proteins in the liver, muscles and cerebrum of 30-day-old rats after labelling with [3H]uridine, [14C]uridine, [3H]cytidine or [3H]orotic acid was measured. It was found that after administration of [3H]uridine, the proteins were 5 - 10 times more radioactive than the RNA. After administration of [14C]uridine, the proteins were 1 - 2 times more heavily labelled than the RNA. Hydrolysis of the proteins followed by chromatography of the amino acids revealed that the protein labelling was mostly due to [3H]glutamate. In the liver, [3H]orotic acid produced very specific labelling of the RNA. The radioactivity of the proteins is very slight. However, the specific labelling of the RNA in the muscles and cerebrum is not so pronounced with this precursor. [3H]Cytidine is an ideal precursor for RNA. The labelling of protein in all three organs examined is very slight, and furthermore, the specific activity of the RNA is 10 - 20 times higher than after labelling with uridine. We were also able to show that after labelling with radioactive uridine, the method of isolation of RNA by alkaline hydrolysis gives incorrect results, because [3H]amino acids interfere with the measurement of the specific activity of the RNA. The heavy labelling of proteins by [3H]-uridine must also be taken into account in histoautoradiography, because our experiments showed that in liver, the proteins in the cell nucleus are 3 times as radioactive as the nucleic acids. The particulate components of the cytoplasm are even 20 times more radioactive than the nucleic acids.
测定了用[³H]尿苷、[¹⁴C]尿苷、[³H]胞苷或[³H]乳清酸标记后30日龄大鼠肝脏、肌肉和大脑中RNA、DNA和蛋白质的放射性。结果发现,给予[³H]尿苷后,蛋白质的放射性比RNA高5 - 10倍。给予[¹⁴C]尿苷后,蛋白质的标记程度比RNA高1 - 2倍。蛋白质水解后进行氨基酸色谱分析表明,蛋白质标记主要是由于[³H]谷氨酸。在肝脏中,[³H]乳清酸对RNA产生非常特异性的标记。蛋白质的放射性非常低。然而,用这种前体对肌肉和大脑中的RNA进行的特异性标记并不那么明显。[³H]胞苷是RNA的理想前体。在所检测的所有三个器官中,蛋白质的标记都非常轻微,此外,RNA的比活性比用尿苷标记后高10 - 20倍。我们还能够表明,用放射性尿苷标记后,通过碱性水解分离RNA的方法会得出错误的结果,因为[³H]氨基酸会干扰RNA比活性的测量。在组织放射自显影中也必须考虑[³H]尿苷对蛋白质的大量标记,因为我们的实验表明,在肝脏中,细胞核中的蛋白质放射性是核酸的3倍。细胞质的颗粒成分的放射性甚至比核酸高20倍。