ANTIGENIC DETERMINANTS OF VARICELLA VIRUS

描述（改编自申请人的摘要）：水痘-带状疱疹病毒 (VZV)是一种进化上古老的α疱疹病毒，具有125 kbp的基因组。 VZV引起两种疾病-水痘和带状疱疹;后一种疾病是一种 VZV向神经性和从病毒中重新激活的显着例证 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 在形成Fc的两种独特的短糖蛋白gE和gI中， 受体复合物 已知疱疹病毒gE/gI复合物是一种重要的 病毒外出和细胞间传播的决定因素，但其机制是 没有很好地理解。 研究计划包含三个具体目标。 要求1 包括对磷酸化和分选基序的表征， gE的C尾。 糖蛋白受体被丝氨酸和丝氨酸修饰， 酪氨酸蛋白激酶;酪氨酸磷酸化只发生在二聚体上。 它是一种gE，以前没有被发现。 磷酸化将是 通过体内和体外蛋白激酶测定来测量。 的事实 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一假设， VZVgE/gI形成多能受体复合物。 目标2寻求通过 诱变方法，以进一步鉴定丝氨酸蛋白激酶， 磷酸化C-尾中不寻常的丝氨酸-脯氨酸-脯氨酸序列， gI，并且还鉴定接近于Gl的内化基序。 磷酸化位点。 在目标3中， 在诱变前后分析gE/gI复合物， 确定每个信号基序对细胞功能的具体作用， 整个复杂。 VZV gE/gI复合物的内吞作用和再循环将被抑制。 详细调查。 作为瞬态的补充策略， 转染试验，重组VZV gE-伪狂犬病病毒将被 在嗜神经性的动物模型中产生和评价，以及VZV突变体 将通过几种成像技术研究gI无效表型， 包括激光扫描共聚焦显微镜和电子显微镜， 免疫标记。 总之，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，表明磷酸化-去磷酸化事件在 参与病毒外出和细胞到细胞的运输和内吞途径 传播.

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.