The UL13 and US3 Protein Kinases of Herpes Simplex Virus 1 Cooperate to Promote the Assembly and Release of Mature, Infectious Virions.
The UL13 and US3 Protein Kinases of Herpes Simplex Virus 1 Cooperate to Promote the Assembly and Release of Mature, Infectious Virions.
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DOI：
10.1371/journal.pone.0131420
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Gershburg E
中科院分区：
文献类型：
--
作者：
Gershburg S;Geltz J;Peterson KE;Halford WP;Gershburg E
1 型单纯疱疹病毒 (HSV-1) 编码两种真正的丝氨酸/苏氨酸蛋白激酶，即 US3 和 UL13 基因产物。 HSV-1 ΔUS3 突变体在培养细胞中以野生型效率复制，HSV-1 ΔUL13 突变体表现出感染性病毒滴度降低 <10 倍。鉴于这些适度的表型，US3 和 UL13 蛋白激酶如何促进 HSV-1 复制仍不清楚。在当前的研究中，我们设计了一组 HSV-1 突变体，其中 UL13 和 US3 基因的部分分别被编码 mCherry 蛋白或绿色荧光蛋白 (GFP) 的表达盒取代，并分析了这些突变体在几种细胞类型中的 DNA 复制、蛋白表达和扩散。仅 US3 功能丧失对病毒 DNA 积累、基因表达、病毒颗粒释放和传播的影响基本上可以忽略不计。单独丧失 UL13 功能也对病毒 DNA 水平没有明显影响。然而，UL13 功能的丧失确实导致两种病毒糖蛋白（gC 和 gD）的稳态水平明显下降、总病毒粒子和感染性病毒粒子的释放以及病毒传播。这两个基因的破坏不会影响病毒 DNA 的积累，但会导致 gC 和 gD 稳态水平进一步降低，并减弱病毒传播和感染性病毒粒子释放。这些数据表明，UL13 激酶在 HSV-1 感染的后期发挥着重要作用，可能是通过影响病毒粒子的组装和/或释放来实现的。此外，数据表明，US3 和 UL13 蛋白激酶的联合活性对于 HSV-1 感染细胞中感染性病毒粒子的有效组装和释放至关重要。
Herpes simplex virus type 1 (HSV-1) encodes two bona fide serine/threonine protein kinases, the US3 and UL13 gene products. HSV-1 ΔUS3 mutants replicate with wild-type efficiency in cultured cells, and HSV-1 ΔUL13 mutants exhibit <10-fold reduction in infectious viral titers. Given these modest phenotypes, it remains unclear how the US3 and UL13 protein kinases contribute to HSV-1 replication. In the current study, we designed a panel of HSV-1 mutants, in which portions of UL13 and US3 genes were replaced by expression cassettes encoding mCherry protein or green fluorescent protein (GFP), respectively, and analyzed DNA replication, protein expression, and spread of these mutants in several cell types. Loss of US3 function alone had largely negligible effect on viral DNA accumulation, gene expression, virion release, and spread. Loss of UL13 function alone also had no appreciable effects on viral DNA levels. However, loss of UL13 function did result in a measurable decrease in the steady-state levels of two viral glycoproteins (gC and gD), release of total and infectious virions, and viral spread. Disruption of both genes did not affect the accumulation of viral DNA, but resulted in further reduction in gC and gD steady-state levels, and attenuation of viral spread and infectious virion release. These data show that the UL13 kinase plays an important role in the late phase of HSV-1 infection, likely by affecting virion assembly and/or release. Moreover, the data suggest that the combined activities of the US3 and UL13 protein kinases are critical to the efficient assembly and release of infectious virions from HSV-1-infected cells.
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