Melvin W T, Milne H B, Slater A A, Allen H J, Keir H M
Eur J Biochem. 1978 Dec;92(2):373-9. doi: 10.1111/j.1432-1033.1978.tb12756.x.
Isolation of newly synthesised RNA can be achieved by treatment of cells in culture with 6-thioguanosine or 4-thiouridine followed by separation of thiol-containing RNA by affinity chromatography on mercurated cellulose columns. After short periods of treatment with 6-thioguanosine the proportion of RNA retained on mercurated cellulose is the same for both poly(A)-containing and poly(A)-free RNA, indicating similar incorporation of the drug into mRNA and rRNA. However, after longer periods of exposure, the cytotoxic effect of 6-thioguanosine results in diminished incorporation of radioactive uridine into RNA and of radioactive leucine into protein; this suggests that synthesis of both RNA and protein are impaired. On the other hand, even after long exposure to high concentrations of 4-thiouridine, the syntheses of RNA and protein are not significantly affected. Proteins synthesised after treatment of cells with 6-thioguanosine are less stable than proteins synthesised after treatment of cells with 4-thiouridine.
新合成RNA的分离可通过用6-硫代鸟苷或4-硫尿苷处理培养的细胞，然后在汞化纤维素柱上通过亲和色谱法分离含硫醇的RNA来实现。用6-硫代鸟苷短期处理后，汞化纤维素上保留的RNA比例对于含聚腺苷酸（poly(A)）和不含聚腺苷酸的RNA是相同的，这表明药物在mRNA和rRNA中的掺入情况相似。然而，长时间暴露后，6-硫代鸟苷的细胞毒性作用导致放射性尿苷掺入RNA以及放射性亮氨酸掺入蛋白质的量减少；这表明RNA和蛋白质的合成均受到损害。另一方面，即使长时间暴露于高浓度的4-硫尿苷，RNA和蛋白质的合成也没有受到显著影响。用6-硫代鸟苷处理细胞后合成的蛋白质比用4-硫尿苷处理细胞后合成的蛋白质稳定性更低。
