Colonno R J, Stone H O
Proc Natl Acad Sci U S A. 1975 Jul;72(7):2611-5. doi: 10.1073/pnas.72.7.2611.
Purified Newcastle disease virus contains an enzyme that incorporates the methyl group from S-adenosyl-L-methionine into RNA synthesized in vitro by the virion-associated RNA polymerase (RNA nucleotidyltransferase). Incorporation of radioactivity from S-adenosyl-L-[methyl-3H]methionine was totally dependent upon RNA synthesis. The methylation reaction was completely inhibited by S-adenosyl-L-homocysteine, suggesting the transfer of only the methyl group of S-adenosyl-methionine to RNA products. Velocity sedimentation and hybridization of the in vitro product RNA indicated that both [3H]methyl and [32P]GMP labels resided in single-stranded 18S RNA molecules which were virus specific. Approximately 1 to 2 methyl groups were incorporated per RNA molecule. DEAE-cellulose chromatography of product RNA after alkaline hydrolysis suggested that the 5' terminus was the site of methylation.
纯化的新城疫病毒含有一种酶,该酶可将S-腺苷-L-甲硫氨酸的甲基基团掺入到由病毒体相关RNA聚合酶(RNA核苷酸转移酶)在体外合成的RNA中。从S-腺苷-L-[甲基-³H]甲硫氨酸掺入放射性完全依赖于RNA合成。S-腺苷-L-高半胱氨酸完全抑制甲基化反应,这表明仅将S-腺苷甲硫氨酸的甲基基团转移到RNA产物中。体外产物RNA的速度沉降和杂交表明,[³H]甲基和[³²P]GMP标记都存在于病毒特异性的单链18S RNA分子中。每个RNA分子大约掺入1至2个甲基基团。碱性水解后产物RNA的DEAE-纤维素色谱分析表明,5'末端是甲基化位点。