Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas, USA.
J Virol. 2013 Dec;87(24):13510-9. doi: 10.1128/JVI.02424-13. Epub 2013 Oct 2.
The herpes simplex virus 1 (HSV-1) immediate-early protein, infected cell protein 22 (ICP22), is required for efficient replication in restrictive cells, for virus-induced chaperone-enriched (VICE) domain formation, and for normal expression of a subset of viral late proteins. Additionally, ICP22 is important for optimal acute viral replication in vivo. Previous studies have shown that the US1 gene that encodes ICP22, produces an in-frame, N-terminally truncated form of ICP22, known as US1.5. To date, studies conducted to characterize the functions of ICP22 have not separated its functions from those of US1.5. To determine the individual roles of ICP22 and US1.5, we made viral mutants that express either ICP22 with an M90A mutation in the US1.5 initiation codon (M90A) or US1.5 with three stop codons introduced upstream of the US1.5 start codon (3×stop). Our studies showed that, in contrast to M90A, 3×stop was unable to replicate efficiently in the eyes and trigeminal ganglia of mice during acute infection, to efficiently establish a latent infection, or to induce VICE domain formation and was only mildly reduced in its replication in restrictive HEL-299 cells and murine embryonic fibroblasts (MEFs). Both mutants enhanced the expression of the late viral proteins virion host shutoff (vhs) and glycoprotein C (gC) and inhibited viral gene expression mediated by HSV-1 infected cell protein 0 (ICP0). When we tested our mutants' sensitivity to type I interferon (beta interferon [IFN-β]) in restrictive cells, we noticed that the plating of the ICP22 null (d22) and 3×stop mutants was reduced by the addition of IFN-β. Overall, our data suggest that US1.5 partially complements the functions of ICP22.
单纯疱疹病毒 1(HSV-1)早期蛋白,感染细胞蛋白 22(ICP22),是在限制细胞中进行有效复制、病毒诱导的伴侣蛋白富集(VICE)结构域形成以及病毒晚期蛋白的正常表达所必需的。此外,ICP22 对体内最佳急性病毒复制也很重要。先前的研究表明,编码 ICP22 的 US1 基因产生 ICP22 的一种框架内、N 端截断形式,称为 US1.5。迄今为止,为了表征 ICP22 的功能而进行的研究尚未将其功能与 US1.5 分开。为了确定 ICP22 和 US1.5 的单独作用,我们构建了表达 ICP22 的病毒突变体,该突变体在 US1.5 起始密码子(M90A)处具有 M90A 突变,或在 US1.5 起始密码子上游引入三个终止密码子(3×stop)的 US1.5。我们的研究表明,与 M90A 相反,3×stop 在急性感染期间无法在小鼠的眼睛和三叉神经节中有效复制,无法有效地建立潜伏感染,也无法诱导 VICE 结构域形成,并且其在限制型 HEL-299 细胞和鼠胚胎成纤维细胞(MEFs)中的复制能力仅轻度降低。两种突变体均增强了晚期病毒蛋白衣壳蛋白关闭(vhs)和糖蛋白 C(gC)的表达,并抑制了由 HSV-1 感染细胞蛋白 0(ICP0)介导的病毒基因表达。当我们在限制细胞中测试我们的突变体对 I 型干扰素(β干扰素[IFN-β])的敏感性时,我们注意到 ICP22 缺失(d22)和 3×stop 突变体的铺板在添加 IFN-β 后减少。总体而言,我们的数据表明 US1.5 部分补充了 ICP22 的功能。