Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, 108-8639, Japan.
Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, 108-8639, Japan.
Nat Commun. 2020 Sep 29;11(1):4894. doi: 10.1038/s41467-020-18718-9.
Identification of the complete set of translated genes of viruses is important to understand viral replication and pathogenesis as well as for therapeutic approaches to control viral infection. Here, we use chemical proteomics, integrating bio-orthogonal non-canonical amino acid tagging and high-resolution mass spectrometry, to characterize the newly synthesized herpes simplex virus 1 (HSV-1) proteome in infected cells. In these infected cells, host cellular protein synthesis is shut-off, increasing the chance to preferentially detect viral proteomes. We identify nine previously cryptic orphan protein coding sequences whose translated products are expressed in HSV-1-infected cells. Functional characterization of one identified protein, designated piUL49, shows that it is critical for HSV-1 neurovirulence in vivo by regulating the activity of virally encoded dUTPase, a key enzyme that maintains accurate DNA replication. Our results demonstrate that cryptic orphan protein coding genes of HSV-1, and probably other large DNA viruses, remain to be identified.
鉴定病毒完整的翻译基因对于理解病毒复制和发病机制以及控制病毒感染的治疗方法非常重要。在这里，我们使用化学蛋白质组学，整合生物正交非典型氨基酸标记和高分辨率质谱，来描述感染细胞中单纯疱疹病毒 1（HSV-1）的新合成蛋白质组。在这些被感染的细胞中，宿主细胞的蛋白质合成被关闭，从而增加了优先检测病毒蛋白质组的机会。我们鉴定了九个先前隐藏的孤儿蛋白编码序列，其翻译产物在 HSV-1 感染的细胞中表达。鉴定的一种蛋白质 piUL49 的功能特征表明，它通过调节病毒编码的 dUTPase 的活性，对 HSV-1 在体内的神经毒力至关重要，dUTPase 是维持准确 DNA 复制的关键酶。我们的结果表明，HSV-1 及可能的其他大型 DNA 病毒的隐藏孤儿蛋白编码基因仍有待鉴定。
