Distinct and separate roles for herpesvirus-conserved UL97 kinase in cytomegalovirus DNA synthesis and encapsidation.
Distinct and separate roles for herpesvirus-conserved UL97 kinase in cytomegalovirus DNA synthesis and encapsidation.
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DOI：
10.1073/pnas.98.4.1895
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发表时间：
2001-02
影响因子：
11.1
通讯作者：
D. Wolf;C. T. Courcelle;M. Prichard;E. Mocarski
中科院分区：
文献类型：
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作者：
D. Wolf;C. T. Courcelle;M. Prichard;E. Mocarski
人巨细胞病毒UL97激酶是抗病毒治疗的一个重要靶点，它至少影响病毒复制的两个不同阶段。与野生型病毒相比，UL97缺失突变体表现出早期复制缺陷，使DNA积累减少4- 6倍，以及晚期衣壳成熟缺陷，导致大多数观察到的复制减少100- 1000倍。抗病毒药物2-溴-5,6-二氯-1-（β -d -核糖呋喃基）-苯并咪唑的阻滞释放实验揭示了UL97激酶在衣壳组装中的重要作用。尽管连接DNA中间体到单位长度基因组的切割未受影响，但后代突变病毒成熟被延迟，在释放该片段后，与野生型相比，后代的积累水平显著降低。透射电镜证实了空a样衣壳的异常积累，既不含病毒DNA，也不含内部支架结构，这与突变病毒感染细胞中无法稳定包装DNA一致。UL97在DNA合成和衣壳组装中的功能表明，这种疱疹病毒保守的激酶介导的蛋白磷酸化提高了病毒复制这两个不同阶段的效率。
The human cytomegalovirus UL97 kinase, an important target of antiviral therapy, has an impact on at least two distinct phases of viral replication. Compared with wild-type virus, the UL97 deletion mutant exhibits an early replication defect that reduces DNA accumulation by 4- to 6-fold, as well as a late capsid maturation defect responsible for most of the observed 100- to 1000-fold reduction in replication. Block-release experiments with the antiviral 2-bromo-5,6-dichloro-1-(beta-D-ribofuranosyl)-benzimidazole revealed an important role for UL97 kinase in capsid assembly. Although cleavage of concatemeric DNA intermediates to unit-length genomes remained unaffected, progeny mutant virus maturation was delayed, with accumulation of progeny at significantly reduced levels compared with wild type after release of this block. Transmission electron microscopy confirmed the aberrant accumulation of empty A-like capsids containing neither viral DNA nor an internal scaffold structure, consistent with a failure to stably package DNA in mutant virus-infected cells. The function of UL97 in DNA synthesis as well as capsid assembly suggests that protein phosphorylation mediated by this herpesvirus-conserved kinase increases the efficiency of these two distinct phases of virus replication.