Functions of the C-terminal domain of varicella-zoster virus glycoprotein E in viral replication in vitro and skin and T-cell tropism in vivo
Functions of the C-terminal domain of varicella-zoster virus glycoprotein E in viral replication in vitro and skin and T-cell tropism in vivo
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DOI：
10.1128/jvi.78.22.12406-12415.2004
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发表时间：
2004-11-01
影响因子：
5.4
通讯作者：
Arvin, AM
中科院分区：
文献类型：
--
作者：
Moffat, J;Mo, CJ;Arvin, AM
水痘-带状疱疹病毒(VZV)糖蛋白E(GE)是水痘-带状疱疹病毒复制所必需的。为了进一步分析GE在VZV复制中的作用，对VZV的62-氨基酸(AA)C-末端结构域进行了完全缺失和点突变。在介导GE内吞作用的Yag1(AA 582至585)、将GE靶向反式高尔基体网络(TGN)的AYRV(AA 568至571)以及包含磷酸化基序(AA 588至601)的“酸性簇”SSTT中引入了靶向突变。利用VZV粘粒将Yag1的Y582G、AYRV的Y569A和SSTT的S593A、S595A、T596A、T598A等替换引入病毒基因组。这些实验证明了这些C-末端基序对VZV复制和毒力的贡献是有层次的。Ge C末端缺失和Yag1突变对VZV的体外复制具有致死性。AYRV和SSTT突变与VZV的恢复相容，但AYRV突变导致病毒在体外迅速传播，SSTT突变导致病毒滴度高于亲本Roka株。当Roka-GE-AYRV和Roka-GE-SSTT突变体在SCIDhu小鼠的皮肤和T细胞移植中进行评估时，干扰TGN靶向与显著的减毒有关，特别是在皮肤中，而SSTT突变不改变VZV在体内的感染性。这些结果首次提供了有关VZV糖蛋白的靶向突变如何在体外影响病毒复制以及在体内完整组织微环境中影响VZV在真皮和表皮细胞以及T细胞中的毒力的信息。
Varicella-zoster virus (VZV) glycoprotein E (gE) is essential for VZV replication. To further analyze the functions of gE in VZV replication, a full deletion and point mutations were made in the 62-amino-acid (aa) C-terminal domain. Targeted mutations were introduced in YAGL (aa 582 to 585), which mediates gE endocytosis, AYRV (aa 568 to 571), which targets gE to the trans-Golgi network (TGN), and SSTT, an "acid cluster" comprising a phosphorylation motif (aa 588 to 601). Substitutions Y582G in YAGL, Y569A in AYRV, and S593A, S595A, T596A, and T598A in SSTT were introduced into the viral genome by using VZV cosmids. These experiments demonstrated a hierarchy in the contributions of these C-terminal motifs to VZV replication and virulence. Deletion of the gE C terminus and mutation of YAGL were lethal for VZV replication in vitro. Mutations of AYRV and SSTT were compatible with recovery of VZV, but the AYRV mutation resulted in rapid virus spread in vitro and the SSTT mutation resulted in higher virus titers than were observed for the parental rOka strain. When the rOka-gE-AYRV and rOka-gE-SSTT mutants were evaluated in skin and T-cell xenografts in SCIDhu mice, interference with TGN targeting was associated with substantial attenuation, especially in skin, whereas the SSTT mutation did not alter VZV infectivity in vivo. These results provide the first information about how targeted mutations of this essential VZV glycoprotein affect viral replication in vitro and VZV virulence in dermal and epidermal cells and T cells within intact tissue microenvironments in vivo.