Varicella-zoster virus infection of human foreskin fibroblast cells results in atypical cyclin expression and cyclin-dependent kinase activity
Varicella-zoster virus infection of human foreskin fibroblast cells results in atypical cyclin expression and cyclin-dependent kinase activity
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DOI：
10.1128/jvi.00163-06
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发表时间：
2006-06-01
影响因子：
5.4
通讯作者：
Moffat, Jennifer F.
中科院分区：
文献类型：
--
作者：
Leisenfelder, Stacey A.;Moffat, Jennifer F.
在人类感染过程中，水痘带状疱疹病毒 (VZV) 会感染很少分裂的细胞，如真皮成纤维细胞、分化的角质形成细胞、成熟 T 细胞和神经元，这些细胞都不会主动合成 DNA；然而，VZV 能够有效地感染它们并利用它们的机制来复制病毒基因组。我们假设 VZV 通过失调细胞周期蛋白和激酶来改变细胞内环境以有利于病毒复制。细胞周期蛋白依赖性激酶 (CDK) 和细胞周期蛋白在 VZV 感染的汇合成纤维细胞中表现出非常不寻常的特征：CDK1、CDK2、细胞周期蛋白 B1、细胞周期蛋白 D3 和细胞周期蛋白 A 蛋白的总量增加，并且 CDK2、CDK4 和细胞周期蛋白 B1 的激酶活性同时强烈诱导。细胞周期蛋白 B1 和 D3 早在感染后 24 小时就增加，与 VZV 蛋白合成同时发生。共聚焦显微镜表明，细胞周期蛋白 D3 过度表达仅限于 IE62 产生区域，而细胞周期蛋白 B1 表达在 VZV 斑块上不规则。 CDK 的下游底物，包括 pRb、p107 和 GM130，通过免疫印迹未显示磷酸化，并且感染后 p21 和 p27 蛋白水平增加。最后，尽管细胞周期蛋白表达和高 CDK 活性的互补表明细胞周期已进入 S 和 G(2) 阶段，但 DNA 染色和流式细胞术表明受感染细胞中可能存在 G(1)/S 阻断。这些数据支持早期研究表明药理学 CDK 抑制剂可以抑制培养细胞中 VZV 的复制。
In its course of human infection, varicella-zoster virus (VZV) infects rarely dividing cells such as dermal fibroblasts, differentiated keratinocytes, mature T cells, and neurons, none of which are actively synthesizing DNA; however, VZV is able to productively infect them and use their machinery to replicate the viral genome. We hypothesized that VZV alters the intracellular environment to favor viral replication by dysregulating cell cycle proteins and kinases. Cyclin-dependent kinases (CDKs) and cyclins displayed a highly unusual profile in VZV-infected confluent fibroblasts: total amounts of CDK1, CDK2, cyclin B1, cyclin D3, and cyclin A protein increased, and kinase activities of CDK2, CDK4, and cyclin B1 were strongly and simultaneously induced. Cyclins B1 and D3 increased as early as 24 h after infection, concurrent with VZV protein synthesis. Confocal microscopy indicated that cyclin D3 overexpression was limited to areas of IE62 production, whereas cyclin B1 expression was irregular across the VZV plaque. Downstream substrates of CDKs, including pRb, p107, and GM130, did not show phosphorylation by immunoblotting, and p21 and p27 protein levels were increased following infection. Finally, although the complement of cyclin expression and high CDK activity indicated a progression through the S and G(2) phases of the cell cycle, DNA staining and flow cytometry indicated a possible G(1)/S blockade in infected cells. These data support earlier studies showing that pharmacological CDK inhibitors can inhibit VZV replication in cultured cells.