Miura K, Watanabe K, Sugiura M, Shatkin A J
Proc Natl Acad Sci U S A. 1974 Oct;71(10):3979-83. doi: 10.1073/pnas.71.10.3979.
The 5'-terminal nucleotide sequences of human reovirus double-stranded RNA were determined after labeling the RNA with [(32)P]phosphate by polynucleotide kinase. The 5' terminal were labeled to only a limited extent prior to sequential oxidation, beta-elimination, and phosphomonoesterase treatment, indicating that the terminal phosphates were in a modified, blocked configuration. Each genome segment, after removing the blocking group, contained the same two 5'-terminal sequences: GpApUp in one chain and G()pCp in the other. G()p is a derivative of guanylic acid, probably 2'-O-methyl-Gp, which renders the 5'-terminal sequence resistant to hydrolysis by alkali. The results indicate that the transcription of reovirus double-stranded RNA strats from the 3' end complementary to the G()pCp-terminal, resulting in the synthesis of single-stranded mRNA carrying the same 5' sequence as the G()pCp-chain. The presence of a modified nucleotide at the 5' terminus of the strand complementary to the mRNA template is a feature common to another double-stranded RNA virus, cytoplasmic polyhedrosis virus.
用人呼肠孤病毒双链RNA的5′末端核苷酸序列,是在用多核苷酸激酶将RNA用[(32)P]磷酸盐标记后测定的。在进行顺序氧化、β-消除和磷酸单酯酶处理之前,5′末端仅被有限程度地标记,这表明末端磷酸盐处于修饰的、封闭的构型。在去除封闭基团后,每个基因组片段都含有相同的两个5′末端序列:一条链上是GpApUp,另一条链上是G()pCp。G()p是鸟苷酸的衍生物,可能是2′-O-甲基-Gp,它使5′末端序列对碱水解具有抗性。结果表明,呼肠孤病毒双链RNA的转录从与G()pCp末端互补的3′端开始,导致合成携带与G()pCp链相同5′序列的单链mRNA。与mRNA模板互补的链的5′末端存在修饰核苷酸是另一种双链RNA病毒——细胞质多角体病毒共有的特征。