Gedamu L, Dixon G H, Davies P L
Biochemistry. 1977 Apr 5;16(7):1383-91. doi: 10.1021/bi00626a023.
Treatment of rainbow trout testis polyribosomes with ethylenediaminetetraacetic acid released polyadenylated protamine messenger RNA in the form of a ribonucleoprotein (mRNP) particle. This mRNP particle which sedimented at 12-14 S could be identified by hybridization to [3H]poly(U) and was partially purified by two successive sucrose gradient sedimentations. When RNA was extracted from the mRNP particle and used as a template in the wheat germ cell-free protein synthesizing system the sole product of translation was protamine. Of this RNA, 30% contained poly(A) sequences and was shown to comigrate with polyadenylated protamine messenger RNA during polyacrylamide gel electrophoresis. When the proteins of the mRNP particle were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the two prominent polypeptides with apparent molecular weights of 73 000 and 29 000 appeared reproducibly. Treatment of trout testis polyribosomes with puromycin in the presence of 0.5 M KCl released a smaller (8-10S) mRNP particle which was similarly shown to contain protamine messenger RNA. Trout testis postribosomal supernatant fraction possessed 16-18S mRNP particles containing polyadenylated RNA which cosedimented with protamine messenger RNA when the particles were dissociated with sodium dodecyl sulfate.
用乙二胺四乙酸处理虹鳟鱼睾丸多核糖体，以核糖核蛋白（mRNP）颗粒的形式释放出聚腺苷酸化的鱼精蛋白信使核糖核酸。这种沉降系数为12 - 14S的mRNP颗粒可通过与[3H]聚尿苷酸杂交来鉴定，并通过连续两次蔗糖梯度沉降进行部分纯化。当从mRNP颗粒中提取RNA并用作麦胚无细胞蛋白质合成系统中的模板时，唯一的翻译产物是鱼精蛋白。这种RNA中，30%含有聚腺苷酸序列，并且在聚丙烯酰胺凝胶电泳过程中显示与聚腺苷酸化的鱼精蛋白信使核糖核酸一起迁移。当通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳检测mRNP颗粒的蛋白质时，可重复性地出现两条明显的多肽，其表观分子量分别为73000和29000。在0.5M氯化钾存在下用嘌呤霉素处理鳟鱼睾丸多核糖体，释放出一种较小的（8 - 10S）mRNP颗粒，同样显示含有鱼精蛋白信使核糖核酸。鳟鱼睾丸核糖体后上清液部分含有16 - 18S的mRNP颗粒，其含有聚腺苷酸化RNA，当颗粒用十二烷基硫酸钠解离时，这些颗粒与鱼精蛋白信使核糖核酸一起沉降。
