VIRUS ENTRY AS A DETERMINANT OF MOUSE HEPATITIS VIRUS NEUROTROPISM

本项目的总体目标是了解分子 MHC神经发病机制。我们的主要目标是了解 病毒组织和细胞嗜性的分子基础，特别是 关于病毒进入过程的机制和特异性。四 该项目的具体目标如下： 1.研究受体特异性的分子基础 利用MHV。我们将确定受体的序列 病毒受体特异性分子 交互.我们将测量病毒受体的亲和力 交互.我们将测量病毒受体的亲和力 相互作用，以确定结合亲和力是否决定 病毒受体相互作用的特异性。 2.为了表征病毒受体后的分子事件， 约束力我们将探讨钙激活的意义- 依赖性磷脂酶A-2（PLA 2）在病毒进入过程中的作用。我 实验室已经表明，这种酶是瞬时激活的， 在病毒与靶细胞结合后立即进行。我们将研究 病毒是否需要进入以及受体的哪个结构域 分子参与其激活。最后，我们将调查 无论是与内吞作用还是病毒-细胞融合有关。 3.鉴定和表征病毒的其他细胞因子 入境我们将使用噬菌体展示文库筛选方法来鉴定 病毒进入所需的细胞因子我们还将使用 另一种方法，cDNA文库转染/感染方法， 以补充噬菌体展示文库技术。 4.为了产生含有不同S蛋白的假型病毒，使用 MHV缺陷型干扰（DI）RNA作为表达载体。这些 病毒将被用来研究S蛋白在 病毒性神经发病机制

The overall objective of this project is to understand the molecular mechanism of MHC neuropathogenesis. Our major goal is to understand the molecular basis of viral tissue and cellular tropism, particularly concerning the mechanism and specificity of viral entry process. The four specific aims for this project are as follows: 1. To study the molecular basis of the specificity of the receptor utilization by MHV. We will determine the sequence of the receptor molecules responsible for the specificity of the virus-receptor interactions. We will measure the affinity of the virus-receptor interactions. We will measure the affinity of the virus-receptor interactions to determine whether the binding affinity determines the specificity of the virus receptor-interaction. 2. To characterize the molecular events subsequent to virus-receptor binding. We will explore the significance of the activation of calcium- dependent phospholipase A-2 (PLA2) in the virus entry process. My laboratory has shown that this enzyme is activated transiently and immediately following virus binding to the target cells. We will study whether it is necessary for virus entry and which domain of the receptor molecule is involved in its activation. Finally, we will investigate whether it is associated with endocytosis or virus-cell fusion. 3. To identify and characterize additional cellular factors for virus entry. We will use a phage display library screening method to identify the possible cellular factors required for virus entry. We will also use an alternative approach, a cDNA library transfection/infection procedure, to complement the phage display library technique. 4. To generate pseudotype viruses containing different S proteins, using the MHV-defective-interfering (DI) RNA as a expression vector. These viruses will be used to study the potential function of the S protein in viral neuropathogenesis.