Greenberg J R
Nucleic Acids Res. 1980 Dec 11;8(23):5685-701. doi: 10.1093/nar/8.23.5685.
A set of proteins crosslinked to L-cell mRNA by irradiating polyribosomes with 254 nm ultraviolet light has been identified. 35S-methionine-labeled crosslinked mRNA-protein complexes were isolated by chromatography on oligo(dT)-cellulose under conditions that prevented non-covalent binding of proteins to RNA and to the column. After enzymatic removal of the RNA the proteins were analyzed in sodium dodecylsulfate/polyacrylamide gels. Six proteins having molecular weights of 98,000, 78,000, 75,000, 68,000, 62,000, and 52,000 were crosslinked to mRNA whether intact polyribosomes or EDTA- released mRNA-protein complexes were irradiated. Digestions with specific RNAases and chromatography on oligo(dT)-cellulose were used to show that a protein of 78,000 daltons was the only one crosslinked to poly(A), and the other proteins were crosslinked to sequences other than poly(A). However, a 78,000 dalton protein was also crosslinked to a sequence other than poly(A).
通过用254nm紫外线照射多核糖体，已鉴定出一组与L细胞mRNA交联的蛋白质。在防止蛋白质与RNA及柱子发生非共价结合的条件下，通过寡聚（dT）-纤维素柱层析分离出35S-甲硫氨酸标记的交联mRNA-蛋白质复合物。用酶去除RNA后，在十二烷基硫酸钠/聚丙烯酰胺凝胶中分析蛋白质。无论照射完整的多核糖体还是经乙二胺四乙酸（EDTA）释放的mRNA-蛋白质复合物，均有分子量分别为98,000、78,000、75,000、68,000、62,000和52,000的六种蛋白质与mRNA交联。用特异性核糖核酸酶消化并通过寡聚（dT）-纤维素柱层析表明，78,000道尔顿的蛋白质是唯一与多聚腺苷酸（poly(A)）交联的蛋白质，其他蛋白质与poly(A)以外的序列交联。然而，78,000道尔顿的蛋白质也与poly(A)以外的序列交联。
