Structural Analysis of the HSV-1 ICP27 protein

描述（由申请方提供）：I型单纯疱疹病毒（HSV-1）感染多种动物、组织和培养细胞，并在宿主感觉神经元中建立终身潜伏感染。HSV-1在感染过程中能够完全超越宿主细胞基因表达机制进行自身复制。HSV-1 ICP 27蛋白是感染细胞内该基因表达转化的重要介质。ICP 27结合HSV-1 mRNA和几种病毒和细胞蛋白以促进病毒基因表达。然而，这些相互作用所需的ICP 27区域已被广泛绘制，并且没有ICP 27的结构信息。目的1的目的是使用NMR分析确定预测的ICP 27 C-末端蛋白结合结构域的结构。在目的2中，ICP 27结构基序将被特异性突变以改变每个结构域的结构并确定在HSV-1期间对ICP 27功能的影响。在目标3中，ICP 27蛋白复合物将被分离和纯化，并通过质谱分析，以确定新的蛋白质相互作用。本研究将阐明ICP 27功能结构域的结构，并绘制其在感染过程中的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.