Kish V M, Pederson T
J Biol Chem. 1976 Oct 10;251(19):5888-94.
Polyribosomal messenger RNA from HeLa cells contain 3'-OH-terminal polyadenylate sequences approximately 133 nucleotides in length (weight average). When analyzed at the ribonucleoprotein level of organization these poly(A)-rich sequences are found to contain tightly bound proteins. These proteins remain associated with the poly(A)-rich RNA during affinity chromatography of RNase A and T1-digested polyribosomes on poly(U)-Sepharose in 0.5 M NaCl, and co-elute from the column with the RNA at 50% formamide. Controls establish that the co-purification of the proteins with poly(A) on poly(U)-Sepharose requires the molecular integrity of the poly(A). Polyacrylamide gel electrophoresis resolves the poly(A)-specific proteins into two components of 74,000 and 62,000 molecular weight. The larger protein is the same size as that previously reported to be associated with poly(A)-rich sequences in HeLa heterogeneous nuclear RNA (Kish, V.M., and Pederson, T. (1975), J. Mol. Biol. 95, 227-238). It is concluded that both HeLa nuclear and polyribosomal poly(A) sequences have a protein (62,000 molecular weight) associated with poly(A) appears to be confined only to messenger RNA.
来自HeLa细胞的多核糖体信使RNA含有长度约为133个核苷酸(重均)的3'-OH末端聚腺苷酸序列。当在核糖核蛋白组织水平进行分析时,发现这些富含聚腺苷酸的序列含有紧密结合的蛋白质。在0.5M NaCl中,用聚(U)-琼脂糖对核糖核酸酶A和T1消化的多核糖体进行亲和层析时,这些蛋白质与富含聚腺苷酸的RNA保持结合,并在50%甲酰胺条件下与RNA一起从柱上共同洗脱。对照实验表明,蛋白质与聚(U)-琼脂糖上的聚腺苷酸共纯化需要聚腺苷酸的分子完整性。聚丙烯酰胺凝胶电泳将聚腺苷酸特异性蛋白质分离为分子量分别为74,000和62,000的两个组分。较大的蛋白质与先前报道的与HeLa异质核RNA中富含聚腺苷酸的序列相关的蛋白质大小相同(基什,V.M.,和佩德森,T.(1975年),《分子生物学杂志》95,227 - 238)。得出的结论是,HeLa细胞核和多核糖体的聚腺苷酸序列都有与聚腺苷酸相关的蛋白质(分子量62,000),该蛋白质似乎仅局限于信使RNA。