Ng T I, Talarico C, Burnette T C, Biron K, Roizman B
The Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Illinois 60637, USA.
Virology. 1996 Nov 15;225(2):347-58. doi: 10.1006/viro.1996.0609.
The predicted amino acid sequence of the human cytomegalovirus U(L)97 protein bears partial homology to the herpes simplex virus 1 U(L)13 protein, especially in regions that are homologous to conserved domains characteristic of protein kinases. Earlier studies showed that U(L)13 mediated the posttranslational processing of several herpes simplex virus 1 proteins including ICP22. Whereas no kinase activity has been specifically attributed to U(L)13, it has been shown that U(L)97 can phosphorylate ganciclovir. To examine whether U(L)97 can substitute for U(L)13, we constructed a herpes simplex virus 1 recombinant virus, R4970, in which U(L)13 was replaced by U(L)97 and in addition, the thymidine kinase gene was deleted. Characterization of this recombinant virus showed the following: (1) The recombinant virus grew as well as the wild-type virus in BHKTK+ cells, which restricted the growth of the U(L)13 virus. (2) U(L)97 could partially mediate the posttranslational modification of HSV-1 ICP22. This modification correlated with the restoration of the amounts of ICP0 and U(S)11 proteins, which were down regulated in the U(L)13- virus-infected cells. (3) The recombinant virus was sensitive to ganciclovir in Vero- and KHOS-infected cells but not in the 143 thymidine kinase minus cells derived from KHOS cells. Vero cells infected with this recombinant virus phosphorylated ganciclovir. We conclude that U(L)97 partially compensates for U(L)13 functions.
人巨细胞病毒U(L)97蛋白的预测氨基酸序列与单纯疱疹病毒1型U(L)13蛋白具有部分同源性,尤其是在与蛋白激酶特征性保守结构域同源的区域。早期研究表明,U(L)13介导了包括ICP22在内的几种单纯疱疹病毒1型蛋白的翻译后加工。虽然尚未明确将激酶活性归因于U(L)13,但已表明U(L)97可使更昔洛韦磷酸化。为了研究U(L)97是否能替代U(L)13,我们构建了一种单纯疱疹病毒1型重组病毒R4970,其中U(L)13被U(L)97取代,此外,胸苷激酶基因被删除。对这种重组病毒的特性分析表明:(1) 该重组病毒在BHKTK+细胞中的生长情况与野生型病毒相同,而BHKTK+细胞会限制U(L)13病毒的生长。(2) U(L)97可部分介导HSV-1 ICP22的翻译后修饰。这种修饰与ICP0和U(S)11蛋白量的恢复相关,这两种蛋白在U(L)13基因缺失病毒感染的细胞中表达下调。(3) 该重组病毒在Vero细胞和KHOS细胞感染时对更昔洛韦敏感,但在源自KHOS细胞的143胸苷激酶缺陷细胞中不敏感。感染这种重组病毒的Vero细胞可使更昔洛韦磷酸化。我们得出结论,U(L)97可部分补偿U(L)13的功能。