Structural Analysis of the HSV-1 ICP27 protein

描述(申请人提供)：单纯疱疹病毒I型(HSV-1)感染多种动物、组织和培养细胞，并在宿主的感觉神经元中建立终身潜伏感染。HSV-1具有在感染过程中完全超越宿主细胞基因表达机制进行自身复制的能力。HSV-1 ICP27蛋白是该基因在感染细胞内表达转化的重要介体。ICP27与HSV-1mRNA及多种病毒和细胞蛋白结合，促进病毒基因表达。然而，这些相互作用所需的ICP27所需的区域已被广泛绘制，并且没有ICP27的结构信息。目的1的目的是通过核磁共振分析确定预测的ICP27 C末端蛋白结合域的结构。在目标2中，ICP27结构基序将被特异性突变，以使每个结构域的结构失活，并确定在HSV-1过程中对1CP27功能的影响。在目标3中，ICP27蛋白质复合体将被分离和纯化，并用质谱学进行分析，以确定新的蛋白质相互作用。这项研究将阐明ICP27功能结构域的结构，并绘制其在感染过程中动态相互作用的完整阵列。

DESCRIPTION (provided by applicant): Herpes Simplex virus type I (HSV-1) infects a wide variety of animals, tissues and cultured cells and established life long latent infections in the sensory neurons of the host. HSV-1 has the ability to completely overtake the host cell gene expression machinery for its own replication during infection. The HSV-1 ICP27 protein is an important mediator of this gene expression conversion within the infected cell. ICP27 binds HSV-1 mRNA and several viral and cellular proteins to promote viral gene expression. However, the regions required of ICP27 required for these interactions have been broadly mapped and there is no structural information for ICP27. The purpose of Aim 1 is to determine the structure of the predicted ICP27 C-terminal protein binding domain using NMR analysis. In Aim 2, the ICP27 structural motifs will be specifically mutated to inactivate the structure of each domain and determine the effect on 1CP27 functions during HSV-1. In Aim 3, ICP27 protein complexes will be isolated and purified and analyzed by Mass Spectrometry in order to identify novel protein interactions. This study will elucidate the structure of the ICP27 functional domains and chart the complete array of its dynamic interactions during infection.