Lee S B, Esteban M
Department of Biochemistry, SUNY Health Science Center, Brooklyn 11203.
Virology. 1993 Apr;193(2):1037-41. doi: 10.1006/viro.1993.1223.
The interferon-induced, double-stranded RNA (dsRNA)-activated protein kinase (p68 kinase) has long been implicated as one of the antiviral agents responsible for overcoming virus infections. To investigate the antiviral potential of p68 kinase, we have generated a recombinant vaccinia virus that expresses human p68 kinase under the control of lac operator/repressor element. Upon induction of p68 kinase gene with the inducer isopropyl-beta-D-thiogalactoside (IPTG), we observed in cultured cells a severe (> 90%) inhibition of virus protein synthesis; this inhibition correlated with autophosphorylation of p68 kinase. As a result of inhibition in the synthesis of virus polypeptides, there was a 100-fold decrease in virus yields. When cells were infected with the recombinant virus expressing lys296-->arg296 mutant p68 kinase there was no reduction in virus yields. Our findings demonstrate that human p68 kinase once activated severely inhibits vaccinia virus replication as a result of inhibition of protein synthesis.
长期以来，干扰素诱导的双链RNA（dsRNA）激活蛋白激酶（p68激酶）一直被认为是负责克服病毒感染的抗病毒因子之一。为了研究p68激酶的抗病毒潜力，我们构建了一种重组痘苗病毒，该病毒在乳糖操纵子/阻遏元件的控制下表达人p68激酶。在用异丙基-β-D-硫代半乳糖苷（IPTG）诱导p68激酶基因后，我们在培养细胞中观察到病毒蛋白合成受到严重抑制（>90%）；这种抑制与p68激酶的自磷酸化相关。由于病毒多肽合成受到抑制，病毒产量下降了100倍。当细胞感染表达lys296→arg296突变型p68激酶的重组病毒时，病毒产量没有降低。我们的研究结果表明，人p68激酶一旦被激活，由于蛋白质合成受到抑制，会严重抑制痘苗病毒的复制。
