Proteomics Analysis of Herpes Simplex Virus Type 1-Infected Cells Reveals Dynamic Changes of Viral Protein Expression, Ubiquitylation, and Phosphorylation
Proteomics Analysis of Herpes Simplex Virus Type 1-Infected Cells Reveals Dynamic Changes of Viral Protein Expression, Ubiquitylation, and Phosphorylation
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DOI：
10.1021/pr301157j
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发表时间：
2013-04-01
影响因子：
4.4
通讯作者：
Radtke, Kerstin
中科院分区：
文献类型：
--
作者：
Bell, Christina;Desjardins, Michel;Radtke, Kerstin
疱疹病毒是最复杂和最广泛的人类病毒之一，并引起从唇疱疹到生殖器感染和脑炎的许多疾病。虽然已经研究了病毒颗粒的组成，但对感染细胞中整个病毒蛋白质组的表达知之甚少。在这里，我们分析了蛋白质组的原型单纯疱疹病毒1型（HSV 1）在感染细胞的质谱。使用高灵敏度LTQ-Orbitrap，我们实现了非常高的蛋白覆盖率，并确定了总共67个结构和非结构病毒蛋白。我们还确定了90个新的磷酸化位点和10个新的泛素化位点在不同的病毒蛋白。在9个HSV 1蛋白上观察到泛素化。我们在大约一半的检测到的病毒蛋白质上鉴定出磷酸化位点;许多高度磷酸化的蛋白质已知调节基因表达。用DNA复制抑制剂治疗引起病毒蛋白丰度和修饰的变化，突出了病毒蛋白在生命周期中的相互依赖性。鉴于表达动力学、泛素化和磷酸化对蛋白质功能的重要性，这些发现将成为未来疱疹病毒生物学研究的重要工具。
Herpesviruses are among the most complex and widespread human viruses and cause a number of diseases ranging from cold sores to genital infections and encephalitis. While the composition of viral particles has been studied, less is known about the expression of the whole viral proteome in infected cells. Here, we analyzed the proteome of the prototypical Herpes Simplex Virus type 1 (HSV1) in infected cells by mass spectrometry. Using a high sensitivity LTQ-Orbitrap, we achieved a very high level of protein coverage and identified a total of 67 structural and nonstructural viral proteins. We also identified 90 novel phosphorylation sites and 10 novel ubiquitylation sites on different viral proteins. Ubiquitylation was observed on nine HSV1 proteins. We identified phosphorylation sites on about half of the detected viral proteins; many of the highly phosphorylated ones are known to regulate gene expression. Treatment with inhibitors of DNA replication induced changes of both viral protein abundance and modifications, highlighting the interdependence of viral proteins during the life cycle. Given the importance of expression dynamics, ubiquitylation, and phosphorylation for protein function, these findings will serve as important tools for future studies on herpesvirus biology.