Levy S, Wood P, Grunstein M, Kedes L
Cell. 1975 Mar;4(3):239-48. doi: 10.1016/0092-8674(75)90171-3.
Newly synthesized polysomal messenger RNAs from cleavage stage embryos of the sea urchin Arbacia punctulata and Lytechinus pictus that contain putative histone mRNAs have been fractionated on 6% polyacrylamide slab gels. At least 8 RNA species with unique electrophoretic mobilities have been recognized. The complex of RNAs has been eluted from the gels in three groups, A, B, and C, in increasing order of mobility. The template activity of the three fractions and the unfractionated starting material was examined in the mouse Krebs II ascites tumor cell-free protein synthesizing system. The unfractionated messenger complex programs the synthesis of proteins that co-electrophorese exclusively with sea urchin histones in both sodium dodecyl sulfate and acid urea gel systems. The products of in vitro protein systhesis stimulated by the individual polyacrylamide gel RNA fractions were similarly examined. Each stimulated protein synthesis and was enriched for specific histone templates. We conclude that RNA fraction A is template for histone f1, C is template for histone f2a1, and B serves as template for f2b, f2a2, and f3 histones. A minor degree of contamination of the A and B RNA fractions was obvious from the production of other histones by each template. The co-electrophoresis of specific template activity with specific radiolabeled RNAs supports the concept that most or all of the labeled RNAs are indeed themselves the histone mRNAs.
从海胆刺冠海胆（Arbacia punctulata）和花斑海胆（Lytechinus pictus）卵裂期胚胎中提取的新合成的多聚核糖体信使核糖核酸（mRNA），其中含有假定的组蛋白mRNA，已在6%的聚丙烯酰胺平板凝胶上进行了分级分离。已识别出至少8种具有独特电泳迁移率的RNA种类。RNA复合物已从凝胶中按迁移率递增的顺序洗脱为A、B、C三组。在小鼠克雷布斯II腹水癌细胞无细胞蛋白质合成系统中检测了这三个级分以及未分级的起始材料的模板活性。未分级的信使复合物指导合成的蛋白质，在十二烷基硫酸钠和酸性尿素凝胶系统中，与海胆组蛋白完全共电泳。对由各个聚丙烯酰胺凝胶RNA级分刺激的体外蛋白质合成产物也进行了类似检测。每个级分都刺激了蛋白质合成，并富含特定的组蛋白模板。我们得出结论，RNA级分A是组蛋白f1的模板，C是组蛋白f2a1的模板，B是f2b、f2a2和f3组蛋白的模板。从每个模板产生其他组蛋白可以明显看出，A和B RNA级分存在轻微程度的污染。特定模板活性与特定放射性标记RNA的共电泳支持了这样一种观点，即大多数或所有标记的RNA实际上本身就是组蛋白mRNA。
