VIRUS ENTRY AS A DETERMINANT OF MOUSE HEPATITIS VIRUS NEUROTROPISM

这个项目的总体目标是理解分子 MHC的神经发病机制。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。四个 该项目的具体目标如下： 1.研究受体特异性的分子基础 MHV的利用率。我们将确定受体的序列 负责病毒受体特异性的分子 互动。我们将测量病毒受体的亲和力 互动。我们将测量病毒受体的亲和力 交互，以确定绑定亲和性是否确定 病毒受体-相互作用的特异性。 2.确定病毒受体后的分子事件 有约束力的。我们将探索钙激活的意义- 在病毒进入过程中依赖的磷脂酶A-2(PLA2)。我的 实验室已经表明，这种酶是瞬时激活的， 紧跟在病毒与目标细胞的结合之后。我们会研究 它是否是病毒进入所必需的，以及受体的哪个区域 分子参与了它的激活。最后，我们将调查 无论它与内吞作用或病毒-细胞融合有关。 3.确定和表征病毒的其他细胞因子 进入。我们将使用噬菌体展示文库筛选方法来鉴定 病毒进入所需的可能的细胞因子。我们还将使用 另一种方法是cDNA文库转染/感染程序， 以补充噬菌体展示文库技术。 4.制备含有不同S蛋白的伪型病毒，使用 将MHV缺陷干扰(DI)RNA作为表达载体。这些 病毒将被用于研究S蛋白在体内的潜在功能 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.