ASSEMBLY AND FUNCTION OF THE HERPES SIMPLEX ENVELOPE

单纯疱疹病毒包膜的组装和功能

• 批准号: 2882200
• 负责人: DUNCAN W. WILSON
• 金额: $ 23.07万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2882200
• 关键词: capsid; glycoprotein structure; glycoproteins; herpes simplex virus 1; immunocytochemistry; immunoprecipitation; intracellular transport; laboratory rabbit; membrane fusion; protein transport; site directed mutagenesis; southern blotting; temperature sensitive mutant; tissue /cell culture; virion; virus DNA; virus assembly; virus envelope; virus infection mechanism; virus protein

Herpes viruses are associated with a range of human disease; Burkitts lymphoma and nasopharyngeal carcinoma (Epstein-Barr virus), cytomegalic inclusion disease (Cytomegalovirus), chickenpox and shingles (Varicella- Zoster virus) and oral or genital mucocutaneous lesions (Herpes simplex virus, HSV types 1 and 2). Their ability to remain latent, then reactivate means they present a serious source of infection following immune suppression in organ transplant, chemotherapy of AIDS patients. The recent appearance of acyclovir-resistant HSV strains in CD4-suppressed subjects underlines the importance of developing new therapeutic strategies to combat herpes virus infections. Little is known of how herpes viruses enter and leave cells. Envelope/plasma membrane fusion during infection requires a large number of viral proteins, and the molecular role of each is unknown. Similarly, the mechanism by which newly assembled progeny capsids become enveloped then released from the cell remains unclear. The long term objective of these studies is a molecular dissection of these phenomena, revealing much about the basic biology of the herpes viruses and potentially yielding a wealth of new virus-specific drug targets. The specific aims are as follows: (1). To investigate the kinetics and morphology of HSV-1 egress from infected cells. A single, synchronized wave of egressing virions will be assayed for DNA packaging, envelopment, acquisition of infectivity and rate of secretion from the cell. Antibodies against the viral capsid will be used to correlate these events with morphological phenomena. (2). To determine the origin of the HSV-1 envelope. The essential HSV glycoprotein gH will be targeted to organelles within HSV infected cells to identify the site of envelopment. Passage of virions through subcellular compartments will be monitored and the kinetics of egress determined. (3). To characterize the defect in the gH mutant tsQ26. Pulse chase experiments will be used to test the hypothesis that the tsQ26 allele of gH is unable to traffic to the site of HSV envelopment at the nonpermissive temperature. (4). To identify regions of glycoprotein gH required for fusion. Regions of the glycoprotein required for envelope/plasma membrane fusion and cell/cell fusion will be determined using a transient expression/complementation assay.

疱疹病毒与一系列人类疾病有关;伯基特 淋巴瘤和鼻咽癌（EB病毒），巨细胞性 包涵体病（巨细胞病毒）、水痘和带状疱疹（水痘- 带状疱疹病毒）和口腔或生殖器粘膜皮肤病变（单纯疱疹 病毒，HSV 1型和2型）。 他们保持潜伏然后重新激活的能力 意味着它们在免疫后是一个严重的感染源， 抑制器官移植，艾滋病患者化疗。 近期 CD 4抑制受试者中出现阿昔洛韦耐药HSV株 强调了开发新的治疗策略的重要性， 预防疱疹病毒感染。 人们对疱疹病毒如何进入和离开细胞知之甚少。 感染期间的包膜/质膜融合需要大量的 病毒蛋白质，每一种的分子作用是未知的。 类似地， 新组装的子代衣壳被包裹的机制， 从细胞中释放的物质仍不清楚。 这些长期目标 研究是对这些现象的分子解剖，揭示了许多关于 疱疹病毒的基本生物学， 新的病毒特异性药物靶点。 具体目标如下： （一）. 为了研究HSV-1的动力学和形态， 被感染的细胞 一个单一的，同步的病毒体外流波将是 检测DNA包装、扩增、感染性和感染率 细胞分泌物。 针对病毒衣壳的抗体将被 用于将这些事件与形态学现象相关联。 （二）、 以确定HSV-1病毒包膜的来源。 基本的HSV 糖蛋白gH将靶向HSV感染细胞内的细胞器， 确定安装地点。 病毒粒子通过亚细胞 将监测隔间并确定出口动力学。 （三）、 为了表征gH突变体tsQ 26中的缺陷。 脉冲追踪 将使用实验来检验gH的tsQ 26等位基因 无法在非许可的条件下访问HSV许可的站点 温度 （四）、 鉴定融合所需的糖蛋白gH区域。 区域 包膜/质膜融合所需的糖蛋白， 细胞/细胞融合将使用瞬时 表达/互补测定。