MECHANISM OF PH INDEPENDENT HSV ENTRY

PH 独立 HSV 进入机制

• 批准号: 2597530
• 负责人: A. OVETA FULLER
• 金额: $ 16.82万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2597530
• 关键词: flow cytometry; gene expression; herpes simplex virus 1; protein structure function; virus infection mechanism; virus protein; virus receptors; western blottings

DESCRIPTION: The present proposal will explore the means by which herpes simplex virus type I (HSV-1) enters cells by pH independent mechanisms. The applicant previous discovered a cell line that is refractory to HSV-1 entry, but which can replicate HSV-1 when infectious DNA is transfected. Utilizing transfection with libraries of HeLa cDNA clones, a novel human gene, B5 was discovered in the applicant's laboratory. This clone is different from the recently discovered human HVEM gene. The studies outlined here will center on the characterization of this gene and its role as a receptor for viral entry. The specific aims of the project are: 1) to characterize the structure and tissue distribution of B5 and to prepare tools for functional analysis including stably transfected cell lines and antibodies; 2) to determine the function of B5 in HSV entry, spread and infection by assessing stable binding, penetration and un- coating during entry of HSV-1. Investigations will also be performed to determine if B5 influences HSV-1 spread, whether it interacts with ligands, and whether it associates with HVEM; 3) to examine if and how B5 interacts with viral and cellular proteins to mediate HSV entry and membrane fusion.

描述：目前的提案将探讨疱疹病毒 单纯疱疹病毒I型(HSV-1)通过不依赖于pH的机制进入细胞。 申请人之前发现了一种对HSV-1无效的细胞系 进入，但它可以复制HSV-1，当感染的DNA被感染时。 利用HeLa基因克隆文库转染人 基因B5是在申请人的实验室中发现的。这个克隆人是 与最近发现的人类HIV基因不同。这些研究 这里概述的将集中在该基因的特征和其 作为病毒进入的受体。该项目的具体目标是： 1)鉴定B5和TO的结构和组织分布 为功能分析准备工具，包括稳定转染的细胞 品系和抗体；2)为了确定B5在HSV进入中的功能， 通过评估稳定的结合、渗透和非结合来传播和感染 HSV-1进入时的涂层。还将进行调查，以 确定B5是否影响HSV-1传播，是否与 配体，以及它是否与HVEM结合；3)检查B5是否以及如何 与病毒和细胞蛋白相互作用，介导HSV进入和 膜融合。