The U(L)31 and U(L)34 gene products of herpes simplex virus 1 are required for optimal localization of viral glycoproteins D and M to the inner nuclear membranes of infected cells.
The U(L)31 and U(L)34 gene products of herpes simplex virus 1 are required for optimal localization of viral glycoproteins D and M to the inner nuclear membranes of infected cells.
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单纯疱疹病毒 1 的 U(L)31 和 U(L)34 基因产物是病毒糖蛋白 D 和 M 最佳定位到感染细胞内核膜所必需的。
DOI：
10.1128/jvi.02431-08
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发表时间：
2009
影响因子：
5.4
通讯作者：
Baines,JoelD
中科院分区：
文献类型：
--
作者：
Wills,Elizabeth;Mou,Fan;Baines,JoelD
单纯疱疹病毒1型的UL 31和UL 34形成核衣壳在内核膜（INM）处出芽所必需的复合物。先前的免疫金电子显微镜和电子断层扫描检查表明，pUL 31，pUL 34，和糖蛋白D和M被招募到核周病毒粒子和密集染色区域的INM核衣壳芽进入核周空间。我们现在通过定量免疫金电子显微镜结合方差分析表明，在感染UL 34无效病毒的细胞的INM和外核膜（ONM）处，gD特异性免疫反应性显著降低。虽然与核膜（NM）相关的gM的量在用UL 34无效病毒感染的细胞中仅略微减少（P= 0.027），但以ONM中的gM为代价的INM中gM的富集极大地依赖于UL 34（P< 0.0001）。pUL 34还与感染细胞裂解物中未成熟形式的gD（预期存在于内质网或核膜中的种类）以及与兔网织红细胞裂解物中与谷胱甘肽S-转移酶融合的gD的胞质尾部直接或间接相互作用，表明pUL 34/gD相互作用在将gD募集到NM中的作用。通过流式细胞术测定，UL 34对gD和gM定位的影响不是gD和gM总表达降低的结果。单独地，pUL 31被设计用于将gD和gM靶向NM的两个小叶，但是对于（i）感染细胞中gD和gM的适当的INM与ONM比率和（ii）INM中代表核衣壳出芽位点的电子致密区域的存在是必需的。我们的结论是，除了他们的作用，在核衣壳蛋白和纤层改变，UL 31和UL 34发挥单独的，但相关的作用，在招聘适当的组件核衣壳出芽网站在INM。
UL31 and UL34 of herpes simplex virus type 1 form a complex necessary for nucleocapsid budding at the inner nuclear membrane (INM). Previous examination by immunogold electron microscopy and electron tomography showed that pUL31, pUL34, and glycoproteins D and M are recruited to perinuclear virions and densely staining regions of the INM where nucleocapsids bud into the perinuclear space. We now show by quantitative immunogold electron microscopy coupled with analysis of variance that gD-specific immunoreactivity is significantly reduced at both the INM and outer nuclear membrane (ONM) of cells infected with a UL34 null virus. While the amount of gM associated with the nuclear membrane (NM) was only slightly (P= 0.027) reduced in cells infected with the UL34 null virus, enrichment of gM in the INM at the expense of that in the ONM was greatly dependent on UL34 (P< 0.0001). pUL34 also interacted directly or indirectly with immature forms of gD (species expected to reside in the endoplasmic reticulum or nuclear membrane) in lysates of infected cells and with the cytosolic tail of gD fused to glutathioneS-transferase in rabbit reticulocyte lysates, suggesting a role for the pUL34/gD interaction in recruiting gD to the NM. The effects of UL34 on gD and gM localization were not a consequence of decreased total expression of gD and gM, as determined by flow cytometry. Separately, pUL31 was dispensable for targeting gD and gM to the two leaflets of the NM but was required for (i) the proper INM-versus-ONM ratio of gD and gM in infected cells and (ii) the presence of electron-dense regions in the INM, representing nucleocapsid budding sites. We conclude that in addition to their roles in nucleocapsid envelopment and lamina alteration, UL31 and UL34 play separate but related roles in recruiting appropriate components to nucleocapsid budding sites at the INM.