ANTIGENIC DETERMINANTS OF VARICELLA VIRUS

描述(摘自申请者摘要)：水痘-带状疱疹病毒 疱疹病毒(VZV)是一种进化上古老的甲型疱疹病毒，基因组长125kbp。 VZV引起两种疾病--水痘和带状疱疹；后者是一种 说明VZV的神经趋向性和从一个 潜伏期延长。目前提案的目标是增加 在分子水平上理解结构/功能关系 形成Fc的两种独特的短糖蛋白Ge和Gi 受体复合体。已知疱疹病毒GE/GI复合体是一种重要的 病毒扩散和细胞间传播的决定因素，但其机制是 不是很清楚。《研究计划》包含三个具体目标。目标1 包括对磷酸化和排序基序的表征 通用电气的C-Tail。糖蛋白受体被丝氨酸和丝氨酸修饰。 酪氨酸蛋白激酶；酪氨酸磷酸化仅在二聚体上发生 通用电气的形式，以前没有得到承认。磷酸化将是 通过体内和体外的蛋白激酶分析进行测量。事实是， 丝氨酸和酪氨酸磷酸化基序都是 几种哺乳动物细胞表面受体支持这样的假设 VZVgE/Gi形成多潜能受体复合体。目标2通过一个 进一步鉴定丝氨酸蛋白激酶的诱变方法 磷酸化C-尾部不寻常的丝氨酸-脯氨酸-脯氨酸序列 GI，并确定靠近GI的内化基序 磷酸化位点。在目标3中，两个组件之间的相互作用 Ge/Gi复合体将在诱变前后进行分析，以便 确定每个信号基序在功能上的具体作用 整个建筑群。VZV Ge/Gi复合体的内吞作用和循环 详细调查了。作为瞬变的补充策略 经转染检测，重组VZV GE伪狂犬病病毒将被 在嗜神经性动物模型和VZV突变体中进行生产和评估 GI为零的表型将通过几种成像技术进行研究， 包括激光扫描共聚焦显微镜和电子显微镜 免疫标记。综上所述，VZV Ge/Gi的磷酸化修饰 支持与其他非病毒受体的进化联系，同时 时间，提示磷酸化-去磷酸化事件在 病毒外流和细胞间的转运和内吞途径 散开。

DESCRIPTION (Adapted from Applicant's Abstract): Varicella-zoster virus (VZV) is an evolutionarily ancient alphaherpesvirus with a 125 kbp genome. VZV causes two diseases -- chickenpox and shingles; the latter disease is a remarkable illustration of VZV neurotropism and reactivation from a prolonged latency. The goal of the current proposal is an increased understanding at a molecular level of the structure/function relationships of the two unique short glycoproteins called gE and gI which form a Fc receptor complex. The herpesviral gE/gI complex is known to be an important determinant of viral egress and cell-to-cell spread, but the mechanisms are not well understood. The Research Plan contains three Specific Aims. Aim 1 includes a characterization of the phosphorylation and sorting motifs in the C-tail of gE. The glycoprotein receptor is modified by both serine and tyrosine protein kinases; tyrosine phosphorylation occurs only on a dimeric form of gE and has not been previously recognized. Phosphorylation will be measured by both in vivo and in vitro protein kinase assays. The fact that both serine and tyrosine phosphorylation motifs are common features of several mammalian cell surface receptors supports the hypothesis that VZVgE/gI form a pluripotential receptor complex. Aim 2 seeks through a mutagenesis approach to further identify the serine protein kinase which phosphorylates the unusual serine-proline-proline sequence in the C-tail of gI, and also identify internalization motifs in proximity to the phosphorylation site. In Aim 3, the interaction of the two components of the gE/gI complex will be analyzed before and after mutagenesis in order to determine the specific roles of each signaling motif on the function of the entire complex. Endocytosis and recycling of the VZV gE/gI complex will be investigated in detail. As a complementary strategy to transient transfection assays, recombinant VZV gE-pseudorabies viruses will be produced and evaluated in an animal model of neurotropism, and VZV mutants with a gI null phenotype will be investigated by several imaging techniques, including laser scanning confocal microscopy and electron microscopy with immunolabeling. In summary, the phosphorylation modifications of VZV gE/gI support an evolutionary link with other nonviral receptors and, at the same time, suggest a role for phosphorylation-dephosphorylation events in trafficking and endocytic pathways involved in viral egress and cell-to-cell spread.