A herpes simplex virus scaffold peptide that binds the portal vertex inhibits early steps in viral replication.
A herpes simplex virus scaffold peptide that binds the portal vertex inhibits early steps in viral replication.
复制标题
结合门顶点的单纯疱疹病毒支架肽抑制病毒复制的早期步骤。
DOI：
10.1128/jvi.00421-13
复制
发表时间：
2013
影响因子：
5.4
通讯作者：
Baines,JoelD
中科院分区：
文献类型：
--
作者：
Yang,Kui;Wills,Elizabeth;Baines,JoelD
先前的实验鉴定了一种12-氨基酸（aa）肽，其足以与单纯疱疹病毒1（HSV-1）门户蛋白相互作用，并且是将门户并入衣壳所必需的。在本研究中，细胞在感染后的不同时间进行处理，所述肽由果蝇触角足蛋白的一部分组成，所述部分先前被证明有效地进入细胞，与野生型HSV-1支架肽（YPYYPGEARGAP）或在位置4和5含有变化的对照肽融合。这些4-酪氨酸和5-脯氨酸残基在疱疹病毒支架蛋白中是高度保守的，并且先前被证明对于门户相互作用是关键的。在感染早期用亚毒性水平的野生型肽治疗使病毒感染性降低超过1,000倍，而突变肽对病毒产量几乎没有影响。在野生型肽存在下感染3 h的细胞中，观察到衣壳正常转运到核边缘，如通过荧光显微镜观察到的。然而，通过电子显微镜在薄切片中的观察揭示了与用突变肽处理的感染细胞相比，含DNA衣壳的异常和显著增加。用肽的早期治疗也防止了病毒DNA复制区室的形成。这些数据表明，抗病毒肽在感染早期稳定衣壳，导致DNA保留在其中，并且这种活性与肽结合到门户蛋白相关。这些数据与门静脉顶点是DNA被排出以启动感染的管道的假设一致。
Previous experiments identified a 12-amino-acid (aa) peptide that was sufficient to interact with the herpes simplex virus 1 (HSV-1) portal protein and was necessary to incorporate the portal into capsids. In the present study, cells were treated at various times postinfection with peptides consisting of a portion of the Drosophila antennapedia protein, previously shown to enter cells efficiently, fused to either wild-type HSV-1 scaffold peptide (YPYYPGEARGAP) or a control peptide that contained changes at positions 4 and 5. These 4-tyrosine and 5-proline residues are highly conserved in herpesvirus scaffold proteins and were previously shown to be critical for the portal interaction. Treatment early in infection with subtoxic levels of wild-type peptide reduced viral infectivity by over 1,000-fold, while the mutant peptide had little effect on viral yields. In cells infected for 3 h in the presence of wild-type peptide, capsids were observed to transit to the nuclear rim normally, as viewed by fluorescence microscopy. However, observation by electron microscopy in thin sections revealed an aberrant and significant increase of DNA-containing capsids compared to infected cells treated with the mutant peptide. Early treatment with peptide also prevented formation of viral DNA replication compartments. These data suggest that the antiviral peptide stabilizes capsids early in infection, causing retention of DNA within them, and that this activity correlates with peptide binding to the portal protein. The data are consistent with the hypothesis that the portal vertex is the conduit through which DNA is ejected to initiate infection.