Mizutani T, Tachibana Y
FEBS Lett. 1986 Oct 20;207(1):162-6. doi: 10.1016/0014-5793(86)80032-1.
Suppressor [32P]phosphoseryl-tRNA, prepared using bovine seryl-tRNA synthetase and ATP:seryl-tRNA phosphotransferase, was mixed with rabbit reticulocyte lysates containing endogenous hemoglobin mRNA having the termination codon UGA (opal). The chromatographic pattern of the lysate on Sephacryl S-200 showed that the radioactivity of [32P]phosphate in the hot trichloroacetic acid-precipitate (phosphoprotein) was eluted at the position between mature hemoglobin and globin subunits. The phosphoprotein, obtained by chromatography on S-200, moved to the position corresponding to that of globin readthrough protein on SDS-PAGE. The analyses of the hydrolyzate of the phosphoprotein showed the presence of phosphoserine in the protein. These results suggest that animal opal suppressor tRNA functions in vitro to transfer phosphoserine to the position of the termination codon UGA (opal) on mRNA.
使用牛丝氨酰 - tRNA合成酶和ATP：丝氨酰 - tRNA磷酸转移酶制备的抑制性[32P]磷酸丝氨酰 - tRNA，与含有终止密码子UGA（乳白密码子）的内源性血红蛋白mRNA的兔网织红细胞裂解物混合。裂解物在Sephacryl S - 200上的色谱图谱显示，热三氯乙酸沉淀（磷蛋白）中[32P]磷酸盐的放射性在成熟血红蛋白和珠蛋白亚基之间的位置洗脱。通过在S - 200上进行色谱分离获得的磷蛋白，在SDS - PAGE上迁移到与珠蛋白通读蛋白相对应的位置。磷蛋白水解产物的分析表明该蛋白中存在磷酸丝氨酸。这些结果表明，动物乳白密码子抑制性tRNA在体外发挥功能，将磷酸丝氨酸转移到mRNA上终止密码子UGA（乳白密码子）的位置。
