COVALENT LINKAGE OF A PROTEIN TO A DEFINED NUCLEOTIDE-SEQUENCE AT 5'-TERMINUS OF VIRION AND REPLICATIVE INTERMEDIATE RNAS OF POLIOVIRUS
COVALENT LINKAGE OF A PROTEIN TO A DEFINED NUCLEOTIDE-SEQUENCE AT 5'-TERMINUS OF VIRION AND REPLICATIVE INTERMEDIATE RNAS OF POLIOVIRUS
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DOI：
10.1073/pnas.74.3.961
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发表时间：
1977-01-01
影响因子：
11.1
通讯作者：
BALTIMORE, D
中科院分区：
文献类型：
--
作者：
FLANEGAN, JB;PETTERSSON, RF;BALTIMORE, D
脊髓灰质炎病毒多核糖体RNA的5'末端为pUp。当32p标记的病毒RNA被RNase T1、T2和A完全消化后，在pH为3.5的条件下用纸离子电泳分析，脊髓灰质炎病毒RNA的5”端候选物被回收为向阴极迁移的化合物。用蛋白酶K处理可逆转其迁移方向，表明存在蛋白质。用毒液磷酸二酯酶处理释放了所有的pUp放射性，表明脊髓灰质炎病毒RNA具有蛋白质-pUp 5'末端。用T1 RNase单独处理病毒粒子RNA产生一个蛋白酶k敏感的低核糖核苷酸。利用rna酶A和U2对该寡核苷酸进行分析，显示其结构为蛋白- pu - u -A-A-A-A- c -A- g。对复制中间RNA的消化含有足够的蛋白质- pup，表明这种结构位于大多数新生脊髓灰质炎病毒RNA分子的5'端。一种蛋白质核苷酸结构可能作为启动脊髓灰质炎病毒RNA合成的引物。
The 5''-terminus of poliovirus polyribosomal RNA is pUp. A candidate for the 5''-terminus of poliovirion RNA was recovered as a compound migrating toward the cathode when 32P-labeled virion RNA was completely digested with RNase T1, T2 and A and analyzed by paper ionophoresis at pH 3.5. Treatment with proteinase K reversed its direction of migration, indicating the presence of protein. Treatment with venom phosphodiesterase liberated all of the radioactivity as pUp, suggesting that poliovirion RNA has a protein-pUp 5''-terminus. Treatment of virion RNA with T1 RNase alone generated a proteinase K-sensitive oligoribonucleotide. Analysis of the oligoribonucleotide using RNase A and U2 showed its structure to be protein-pU-U-A-A-A-A-C-A-G. Digests of replicative intermediate RNA contained sufficient protein-pUp to suggest that this structure is at the 5''-end of most nascent poliovirus RNA molecules. A protein-nucleotide structure probably acts as a primer for initiating synthesis of poliovirus RNA.