Early Interactions of DNA Viruses and Host

DNA病毒与宿主的早期相互作用

• 批准号: 6737587
• 负责人: GERD G MAUL
• 金额: $ 31.59万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6737587
• 关键词: Epstein Barr virus; gene induction /repression; herpes simplex virus 1; host organism interaction; nucleic acid sequence; nucleoproteins; reporter genes; simian virus 40; tissue /cell culture; virus DNA; virus genetics; virus infection mechanism; virus replication

DESCRIPTION (provided by applicant): DNA viruses are the causative agents of a large number of debilitating diseases. They are also used as vehicles in gene therapy. We are investigating the early host-virus interaction with the long-term goal of identifying common sites of interference and of prolonging the transcriptional life of non-replicative gene therapy vehicles. DNA viruses are deposited at very few precisely circumscribed nuclear domains named NDI0 suggesting the existence of a targeted DNA deposition mechanism within the nucleus. We mapped the targeting DNA sequence for simian virus 40 (SV40) to the core origin of replication. Also DNA viruses begin to transcribe in association with ND10. Thus, an essential functional interaction appears to reside at ND10. On the other hand the major ND10-associated proteins are interferon-upregulate. We will determine, whether deposition of the viruses at ND10 is essential and beneficial for infection or instead, reduces productive viral infection, and whether the mechanism of deposition is shared by the different DNA virus families that deposit their infectious virus at ND10. Elucidation of such a mechanism could provide general means of interference with viral infections. Therefore, basic biological properties need to be defined. We will continue to define the mechanism that targets viral DNA to ND10 by identifying the DNA targeting sequence using a new method where DNA is directly labeled in vivo. 2. We will define the functional consequences of viral deposition at ND10 by determining whether deposition of a reporter gene at ND10 by the viral DNA targeting sequence determines transcriptional potential. 3. We will determine where quiescent herpes viruses (EBV and HSV-l) transcribe and replicate relative to ND10 in a cell culture model system and whether upon reactivation they start their immediate early transcription at ND10. 4. We will determine whether the absence of certain ND10-associated proteins influences productive infection using knock-out cells and focusing on ND10-associated proteins that are thought to be transcriptional repressors or involved in heterochromatin formation. 5. We will determine whether the SUMO-1 iso-peptidase dependent release of ND10-associated proteins induces or suppresses activation of repressed viral genomes. The long-term goal is to define the mechanism by which the infecting virus deposits DNA at specific highly localized sites within the nucleus, what advantage the virus (or the cell) derives from such specific localization, and to determine whether this specific localization could be used as a target for anti-viral therapy, or has properties to improve long-term transcription of viral vectors for gene therapy.

描述（由申请人提供）：DNA病毒是A 大量衰弱的疾病。它们也被用作基因的车辆 治疗。我们正在研究与早期的宿主病毒相互作用 确定干扰和延长的常见地点的长期目标 非复制基因疗法的转录寿命。 DNA病毒 存放在很少的精确限制的核域名NDI0 表明存在靶向DNA沉积机制 核。我们映射了邻苯二甲酸病毒40（SV40）的靶向DNA序列 复制的核心起源。同样，DNA病毒开始抄录 与ND10。因此，似乎存在于ND10的基本功能相互作用。 另一方面，主要与ND10相关的蛋白是干扰素启用。 我们将确定在ND10上的病毒沉积是否必不可少，并且 有益于感染或减少生产性病毒感染，并 沉积机理是否由不同的DNA病毒共享 将其传染病毒存入ND10的家庭。阐明这样的 机制可以提供一般干扰病毒感染的手段。 因此，需要定义基本的生物学特性。我们将继续 通过识别DNA来定义将病毒DNA靶向ND10的机制 使用新方法靶向序列，其中DNA直接在体内标记。 2。 我们将定义ND10病毒沉积的功能后果 确定病毒DNA在ND10上的沉积是否沉积 靶向序列决定转录电位。 3。我们将确定 静态疱疹病毒（EBV和HSV-L）转录和复制的地方 相对于细胞培养模型系统中的ND10以及是否重新激活 他们从ND10开始立即进行早期转录。 4。我们将确定 缺乏某些ND10相关蛋白会影响生产力 使用基因敲除细胞感染，并专注于ND10相关蛋白 被认为是转录阻遏物或参与异染色质 形成。 5。我们将确定SUMO-1等肽酶是否依赖 ND10相关蛋白的释放会诱导或抑制激活 抑制病毒基因组。 长期目标是定义感染病毒的机制 将DNA沉积在核内的特定高度局部位点，什么 优势病毒（或细胞）来自这种特定的定位，并且 确定该特定本地化是否可以用作 抗病毒疗法，或具有改善长期转录的特性 基因治疗的病毒载体。