Boone R F, Ensinger M J, Moss B
J Virol. 1977 Feb;21(2):475-83. doi: 10.1128/JVI.21.2.475-483.1977.
Guanylyltransferase and methyltransferases that modify the 5'-terminals of viral mRNA's to form the structures m7G(5')pppAm- and m7G(5')pppGm- appear to be synthesized afte- vaccinia virus infection of HeLa cells. Elevations in these enzyme activities were detected within 1 h after virus inoculation and increased 15- to 30-fold by 4 to 10 h. Increases in the guanylyl- and methyltransferase activities were prevented by cycloheximide, an inhibitor of protein synthesis, but not by cytosine arabinoside, an inhibitor of DNA synthesis. The latter results suggest that the mRNA guanylyl- and methyltransferases are "early" or prereplicative viral gene products. The guanylyltransferase and two methyltransferases, a guanine-7-methyltransferase and nucleoside-2'-methyltransferase, were isolated by column chromatography from infected cell extracts and found to have properties similar or identical to those of the corresponding enzyme previously isolated from vaccinia virus cores. In contrast, enzymes with these properties could not be isolated from uninfected cells.
在痘苗病毒感染HeLa细胞后,似乎会合成对病毒mRNA的5'末端进行修饰以形成m7G(5')pppAm-和m7G(5')pppGm-结构的鸟苷酸转移酶和甲基转移酶。在病毒接种后1小时内就检测到这些酶活性升高,到4至10小时时增加了15至30倍。蛋白质合成抑制剂环己酰亚胺可阻止鸟苷酸转移酶和甲基转移酶活性的增加,但DNA合成抑制剂阿糖胞苷则无此作用。后一结果表明,mRNA鸟苷酸转移酶和甲基转移酶是“早期”或复制前的病毒基因产物。通过柱色谱法从感染细胞提取物中分离出鸟苷酸转移酶和两种甲基转移酶,即鸟嘌呤-7-甲基转移酶和核苷-2'-甲基转移酶,发现它们的性质与先前从痘苗病毒核心中分离出的相应酶相似或相同。相比之下,无法从未感染细胞中分离出具有这些性质的酶。