Herpes Simplex Virus Replication: Roles of Viral Proteins and Nucleoporins in Capsid-Nucleus Attachment
Herpes Simplex Virus Replication: Roles of Viral Proteins and Nucleoporins in Capsid-Nucleus Attachment
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DOI：
10.1128/jvi.01139-08
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发表时间：
2009-02-15
影响因子：
5.4
通讯作者：
Brown, Jay C.
中科院分区：
文献类型：
--
作者：
Copeland, Anna Maria;Newcomb, William W.;Brown, Jay C.
单纯疱疹病毒1型（HSV-1）的复制涉及亲本衣壳停靠在宿主核孔复合物（NPC）上的步骤。然后病毒基因组通过核孔易位到核质中，在那里它被转录和复制以传播感染。我们研究了病毒和细胞蛋白在衣壳-核附着过程中的作用。将Vero细胞预先装载对感兴趣的蛋白质具有特异性的抗体，并用含有绿色荧光蛋白标记的衣壳的HSV-1感染，并通过荧光显微镜定量结合到核表面的衣壳。结果显示，核衣壳附着被病毒被膜蛋白VP 1/2（UL 36基因）特异性抗体减弱，但不被UL 37（一种被膜蛋白）、主要衣壳蛋白（VP 5）或VP 23（一种次要衣壳蛋白）特异性类似抗体减弱。用核孔蛋白特异性抗体进行的类似研究证明了Nup 358特异性抗体的减毒作用，但Nup 214不特异性抗体的减毒作用。使用小干扰RNA（siRNA）进一步研究核孔蛋白的作用。在用特异于Nup 214或Nup 358但不特异于TPR的siRNA处理的细胞中，衣壳与细胞核的附着减弱。结果被解释为表明，VP 1/2是参与特定的附着到NPC和/或在迁移的衣壳的核表面。建议衣壳通过Nup 358和Nup 214的复合物附着到NPC，高分辨率免疫荧光研究有利于结合Nup 358。
Replication of herpes simplex virus type 1 (HSV-1) involves a step in which a parental capsid docks onto a host nuclear pore complex (NPC). The viral genome then translocates through the nuclear pore into the nucleoplasm, where it is transcribed and replicated to propagate infection. We investigated the roles of viral and cellular proteins in the process of capsid-nucleus attachment. Vero cells were preloaded with antibodies specific for proteins of interest and infected with HSV-1 containing a green fluorescent protein-labeled capsid, and capsids bound to the nuclear surface were quantified by fluorescence microscopy. Results showed that nuclear capsid attachment was attenuated by antibodies specific for the viral tegument protein VP1/2 (UL36 gene) but not by similar antibodies specific for UL37 (a tegument protein), the major capsid protein (VP5), or VP23 (a minor capsid protein). Similar studies with antibodies specific for nucleoporins demonstrated attenuation by antibodies specific for Nup358 but not Nup214. The role of nucleoporins was further investigated with the use of small interfering RNA (siRNA). Capsid attachment to the nucleus was attenuated in cells treated with siRNA specific for either Nup214 or Nup358 but not TPR. The results are interpreted to suggest that VP1/2 is involved in specific attachment to the NPC and/or in migration of capsids to the nuclear surface. Capsids are suggested to attach to the NPC by way of the complex of Nup358 and Nup214, with high-resolution immunofluorescence studies favoring binding to Nup358.