Katze M G, Persson H, Johansson B M, Philipson L
J Virol. 1983 Apr;46(1):50-9. doi: 10.1128/JVI.46.1.50-59.1983.
Adenovirus type 5 (Ad5) host range mutants dl312 and hr-1, with lesions in region E1A (0 to 4.5 map units) of the viral genome, fail to accumulate virus-specific early RNA during infection in HeLa cells. In a recent report, we showed that the addition of anisomycin, a stringent inhibitor of protein synthesis, at 1 h after infection of HeLa cells with hr-1 virus resulted in the accumulation of properly spliced and translatable mRNA from all early regions (M. G. Katze, H. Persson, and L. Philipson, Mol. Cell. Biol. 1:807-813, 1981). Based on these results we proposed a model in which expression of early mutant RNA was achieved through inactivation of a cellular protein normally causing a reduction in the amount of viral RNA. These studies have been extended in the present report, which shows that early viral proteins can be detected in Ad5 dl312- and Ad5 hr-1-infected HeLa cells which have been treated for several hours with anisomycin either shortly after infection or before infection. A pulse of drug treatment also resulted in expression of substantial amounts of adenovirus structural proteins after infection with both Ad5 hr-1 and Ad5 dl312, whereas in drug-free controls no late proteins were detected. The Ad5 hr-1 virus previously reported to be DNA replication negative in nonpermissive HeLa cells was found to replicate its DNA, albeit at low levels, when anisomycin was present either from 1 to 5 h postinfection or for 5 h before infection. When infectious virus production was examined in mutant-infected cells the titer of Ad5 dl312 virus was found to increase at least 500-fold in anisomycin-treated HeLa cells. Taken together, these and our previous results suggest that the block in gene expression characteristic for complementation group I Ad5 host range mutants in HeLa cells can be overcome by inactivating cellular gene products serving as negative regulators of viral gene expression.
腺病毒5型（Ad5）宿主范围突变体dl312和hr-1，其病毒基因组的E1A区域（0至4.5个图谱单位）存在损伤，在感染HeLa细胞期间无法积累病毒特异性早期RNA。在最近的一份报告中，我们表明，在用hr-1病毒感染HeLa细胞1小时后添加茴香霉素（一种严格的蛋白质合成抑制剂），会导致所有早期区域正确剪接和可翻译的mRNA积累（M.G.卡茨、H.佩尔松和L.菲利普松，《分子细胞生物学》1:807 - 813，1981年）。基于这些结果，我们提出了一个模型，其中早期突变RNA的表达是通过使一种通常会导致病毒RNA量减少的细胞蛋白失活来实现的。这些研究在本报告中得到了扩展，本报告表明，在用茴香霉素处理数小时的Ad5 dl312和Ad5 hr-1感染的HeLa细胞中，可以检测到早期病毒蛋白，这些细胞在感染后不久或感染前接受处理。药物处理脉冲也导致在用Ad5 hr-1和Ad5 dl312感染后表达大量腺病毒结构蛋白，而在无药物对照中未检测到晚期蛋白。先前报道在非允许性HeLa细胞中DNA复制阴性的Ad5 hr-1病毒，当在感染后1至5小时或感染前5小时存在茴香霉素时，被发现能够复制其DNA，尽管水平较低。当在突变体感染的细胞中检测传染性病毒产生时，发现Ad5 dl312病毒在经茴香霉素处理的HeLa细胞中的滴度至少增加了500倍。综上所述，这些以及我们之前的结果表明，通过使作为病毒基因表达负调节因子的细胞基因产物失活，可以克服HeLa细胞中I组Ad5宿主范围突变体特有的基因表达阻断。
