Moss B, Keith J M, Gershowitz A, Ritchey M B, Palese P
J Virol. 1978 Jan;25(1):312-8. doi: 10.1128/JVI.25.1.312-318.1978.
Guanylyl- and methyltransferases, isolated from purified vaccinia virus, were used to specifically label the 5' ends of the genome RNAs of influenza A and B viruses. All eight segments were labeled with [alpha-(32)P]guanosine 5'-triphosphate or S-adenosyl[methyl-(3)H]methionine to form "cap" structures of the type m(7)G(5')pppN(m)-, of which unmethylated (p)ppN- represents the original 5' end. Further analyses indicated that m(7)G(5')pppA(m), m(7)G(5')pppA(m)pGp, and m(7)G(5')pppA(m)pGpUp were released from total and individual labeled RNA segments by digestion with nuclease P1, RNase T1, and RNase A, respectively. Consequently, the 5'-terminal sequences of most or all individual genome RNAs of influenza A and B viruses were deduced to be (p)ppApGpUp. The presence of identical sequences at the ends of RNA segments of both types of influenza viruses indicates that they have been specifically conserved during evolution.
从纯化的痘苗病毒中分离出的鸟苷酸转移酶和甲基转移酶,用于特异性标记甲型和乙型流感病毒基因组RNA的5'末端。所有八个片段均用[α-(32)P]鸟苷5'-三磷酸或S-腺苷基[甲基-(3)H]甲硫氨酸进行标记,以形成m(7)G(5')pppN(m)-类型的“帽”结构,其中未甲基化的(p)ppN-代表原始的5'末端。进一步分析表明,通过分别用核酸酶P1、核糖核酸酶T1和核糖核酸酶A消化,m(7)G(5')pppA(m)、m(7)G(5')pppA(m)pGp和m(7)G(5')pppA(m)pGpUp从总标记RNA片段和单个标记RNA片段中释放出来。因此,推断甲型和乙型流感病毒大多数或所有单个基因组RNA的5'-末端序列为(p)ppApGpUp。两种流感病毒RNA片段末端存在相同序列,表明它们在进化过程中得到了特异性保守。