FUNCTIONAL STUDIES OF HERPESVIRUS PROTEINS OF THE VIRION

我们的实验室最近证明， 单纯疱疹病毒1型（HSV-1）的早期蛋白，ICP 4，是局部的 在病毒体的被膜区域内（J. Virol. 63：3338，1989）。 其他 最近的数据表明，ICP 0，而不是ICP 27，也与 纯化的病毒体 相关的初步研究表明，ICP 4是 与病毒感染细胞的质膜部分相关。 基于在纯化的病毒粒子中ICP 4和ICP 0的发现，我们认为， 可能有必要考虑ICP 4和ICP 0可能采取行动的可能性， 与VP 16反式激活蛋白协同作用。 整体 拟议的研究计划的目标是确定的特点， 以及即时早期蛋白质的作用代表了一种新的 关于HSV-1立即早期蛋白的研究方向， 基于关于HSV-1立即早期的新研究方向， 根据现有的初步数据，我们认为， 实验室处于跟踪这些研究的独特位置。 前三个具体目标中的实验旨在定义和 表征ICP 4、ICP 0或ICP 27与病毒体的相互作用， 病毒感染细胞的质膜。 第四个具体目标 重点研究确定病毒体相关蛋白的功能作用 早期蛋白质 在第一个具体目标中，纯化的蛋白质或合成肽 将使用ICP 4、ICP 0和ICP 27制备免疫学试剂。 第二个具体目标的研究将描述病毒体相关的 ICP 4和ICP 0，关于磷酸化和可能的脂肪酸 的增订条文 此外，使用化学交联剂以及无意义的 和缺失突变体将用于进一步表征关联 ICP 4和ICP 0与病毒体的结合。 在第三个具体目标类似 实验将用于表征等离子体之间的相互作用 HSV-1感染细胞的膜和立即早期蛋白。 将检查膜相关ICP 4相对于 磷酸化和脂肪酰基的存在。 废话和 缺失突变体将用于鉴定ICP 4的结构域， 因为它与质膜结合。 第四个具体目标 包括专注于定义病毒体功能作用的研究， 相关的ICP 4和ICP 0。 实验将解决的可能性， 病毒体相关ICP 4和ICP 0作为转录激活因子发挥作用 在病毒复制周期的早期阶段 研究是 还建议确定ICP 4和ICP 0是否代表结构组件 病毒粒子的。

Our laboratory has recently demonstrated that one of the major immediate early proteins of herpes simplex virus type 1 (HSV-1), ICP4, is localized within the tegument region of the virion (J. Virol. 63:3338, 1989). Other more recent data suggest that ICP0, but not ICP27, is also associated with purified virions. Related preliminary studies have shown that ICP4 is associated with the plasma membrane fraction of virus-infected cells. Based on the finding of ICP4 and ICP0 in purified virions, we suggest that it may be necessary to consider the possibility that ICP4 and ICP0 may act synergistically with the VP16 transactivating protein. The overall objective of the proposed research program is to define the characteristics as well as the role of the immediate early proteins represent a new direction of research concerning the HSV-1 immediate early proteins and based on the new direction of research concerning the HSV-1 immediate early proteins and based on the preliminary data available, we believe our laboratory is in a unique position to follow up on these studies. Experiments in the first three specific aims are designed to define and characterize the interaction of ICP4, ICP0 or ICP27 with the virion and the plasma membrane of the virus-infected cell. The fourth specific aim focuses on studies to identify the functional role of the virion-associated immediate early proteins. In the first specific aim, either purified proteins or synthetic peptides of ICP4, ICP0, and ICP27 will be used to prepare immunological reagents. Studies in the second specific aim will characterize the virion-associated ICP4 and ICP0 with regard to phosphorylation and possible fatty acid addition. In addition, use of chemical cross-linkers as well as nonsense and deletion mutants will be used to characterize further the association of ICP4 and ICP0 with the virion. In the third specific aim similar experiments will be used to characterize the interaction between the plasma membrane of HSV-1-infected cells and the immediate early proteins. Membrane-associated ICP4 will be examined relative to the degree of phosphorylation and the presence of fatty acyl groups. Nonsense and deletion mutants will be used to identify the domain of ICP4 responsible for its association with the plasma membrane. The fourth specific aim includes studies focused on defining the functional role of the virion- associated ICP4 and ICP0. Experiments will address the possibility that the virion-associated ICP4 and ICP0 function as transcriptional activators during the very early stages of the virus replication cycle. Studies are also proposed to determine if ICP4 and ICP0 represent structural components of the virion.