Cell Culture Evolution of a Herpes Simplex Virus 1 (HSV-1)/Varicella-Zoster Virus (VZV) UL34/ORF24 Chimeric Virus Reveals Novel Functions for HSV Genes in Capsid Nuclear Egress
Cell Culture Evolution of a Herpes Simplex Virus 1 (HSV-1)/Varicella-Zoster Virus (VZV) UL34/ORF24 Chimeric Virus Reveals Novel Functions for HSV Genes in Capsid Nuclear Egress
复制标题
DOI：
10.1128/jvi.00957-21
复制
发表时间：
2021-06
影响因子：
5.4
通讯作者：
R. Roller;Tineke Hassman;Alison Haugo-Crooks
中科院分区：
文献类型：
--
作者：
R. Roller;Tineke Hassman;Alison Haugo-Crooks
病毒蛋白之间的相互作用对于病毒颗粒的组装和排出至关重要，并且这种相互作用是抗病毒治疗的有吸引力的靶点。关键功能相互作用的识别可能是缓慢而乏味的。摘要单纯疱疹病毒（HSV）和水痘带状疱疹病毒（VZV）同属α疱疹病毒亚科，但分属于不同的属。HSV-1 UL 34编码序列被其VZV同源物开放阅读框24（ORF 24）的编码序列取代，导致病毒在病毒生长、扩散、衣壳排出和核纤层破坏方面具有缺陷，与在UL 34无效病毒中所见的非常相似，尽管ORF 24蛋白和HSV pUL 31之间存在正常的相互作用，并且核排出复合物在核膜处适当定位。对细胞培养物中生长的最小选择导致比亲本嵌合病毒更有效地生长和传播的病毒。这些病毒在支持核纤层破坏、正常核出口复合体定位和衣壳去除的能力上各不相同。将抑制生长缺陷的单突变定位于ORF 24、ICP 22和ICP 4的编码序列，并且一种病毒在ICP 22和US 3编码序列中的每一个中携带单突变。这些病毒的表型支持ICP 22在核纤层破坏中的作用和主要转录调节因子ICP 4在衣壳核出口中的完全出乎意料的作用。病毒蛋白之间的相互作用对于病毒颗粒的组装和排出至关重要，并且这种相互作用是抗病毒治疗的有吸引力的靶点。关键功能相互作用的识别可能是缓慢而乏味的。疱疹病毒的衣壳核排出是装配和排出途径中的关键事件，并且由疱疹病毒中保守的两种蛋白质pUL 31和pUL 34介导。在这里，我们描述了一种细胞培养进化的方法，以确定其他病毒基因产物的功能与pUL 34相互作用。
Interactions among virus proteins are critical for assembly and egress of virus particles, and such interactions are attractive targets for antiviral therapy. Identification of critical functional interactions can be slow and tedious. ABSTRACT Herpes simplex virus (HSV) and varicella-zoster virus (VZV) are both members of the alphaherpesvirus subfamily but belong to different genera. Substitution of the HSV-1 UL34 coding sequence with that of its VZV homolog, open reading frame 24 (ORF24), results in a virus that has defects in viral growth, spread, capsid egress, and nuclear lamina disruption very similar to those seen in a UL34-null virus despite normal interaction between ORF24 protein and HSV pUL31 and proper localization of the nuclear egress complex at the nuclear envelope. Minimal selection for growth in cell culture resulted in viruses that grew and spread much more efficiently that the parental chimeric virus. These viruses varied in their ability to support nuclear lamina disruption, normal nuclear egress complex localization, and capsid de-envelopment. Single mutations that suppress the growth defect were mapped to the coding sequences of ORF24, ICP22, and ICP4, and one virus carried single mutations in each of the ICP22 and US3 coding sequences. The phenotypes of these viruses support a role for ICP22 in nuclear lamina disruption and a completely unexpected role for the major transcriptional regulator, ICP4, in capsid nuclear egress. IMPORTANCE Interactions among virus proteins are critical for assembly and egress of virus particles, and such interactions are attractive targets for antiviral therapy. Identification of critical functional interactions can be slow and tedious. Capsid nuclear egress of herpesviruses is a critical event in the assembly and egress pathway and is mediated by two proteins, pUL31 and pUL34, that are conserved among herpesviruses. Here, we describe a cell culture evolution approach to identify other viral gene products that functionally interact with pUL34.