PROTEINS CROSSLINKED TO MESSENGER-RNA BY IRRADIATING POLYRIBOSOMES WITH ULTRAVIOLET-LIGHT
PROTEINS CROSSLINKED TO MESSENGER-RNA BY IRRADIATING POLYRIBOSOMES WITH ULTRAVIOLET-LIGHT
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DOI：
10.1093/nar/8.23.5685
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发表时间：
1980-01-01
影响因子：
14.9
通讯作者：
GREENBERG, JR
中科院分区：
文献类型：
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作者：
GREENBERG, JR
用254 nm紫外线照射多聚核糖体，可使一组蛋白质与L-细胞mRNA交联。35 S-蛋氨酸标记的交联mRNA-蛋白质复合物在oligo（dT）-纤维素上层析分离，层析条件是防止蛋白质与RNA和层析柱的非共价结合。酶法去除RNA后，在十二烷基硫酸钠/聚丙烯酰胺凝胶中分析蛋白质。分子量为98，000、78，000、75，000、68，000、62，000和52，000的六种蛋白质与mRNA交联，无论是照射完整的多聚核糖体还是EDTA释放的mRNA-蛋白质复合物。使用具有特异性RNA酶的二聚酶和寡聚（dT）-纤维素上的层析来显示78，000道尔顿的蛋白质是唯一与聚（A）交联的蛋白质，并且其他蛋白质与聚（A）以外的序列交联。然而，78，000道尔顿的蛋白质也与聚（A）以外的序列交联。
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).