INTERACTION OF VZV GENES TRANSCRIBED DURING LATENCY IN HUMAN GANGLIA

人类神经节潜伏期转录的 VZV 基因的相互作用

• 批准号: 6410649
• 负责人: RANDALL J. COHRS
• 金额: $ 24.29万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-12-01 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6410649
• 关键词: ganglion cell; genetic mapping; genetic promoter element; genetic transcription; immunoelectron microscopy; latent virus infection; nervous system infection; neurotropic virus; tissue /cell culture; transfection; varicella zoster virus; virus genetics; virus protein; virus replication

Reactivation of latent varicella zoster virus (VZV) from dorsal root ganglia at all levels of the neuraxis produces serious neurological disease. In latently infected ganglia from normal humans, we have shown that low levels of VZV DNA are present in a circular form, that VZV genes 21, 29, 62 and 63 are transcribed, and that VZV gene 63 is translated. The fact that the VZV gene transcribed during latency include those immediate- early (IE) genes with either proven (gene 62) or implied (gene 63) regulatory functions and genes with putative DNA binding capacity (VZV genes 21 and 29) provides the ratione for our hypothesis that proteins encoded by VZV genes transcribed during latency inhibit virus replication. Synthesis of virus proteins and assembly of infectious virions are complex events that depend on coordinated gene activity and differ during productive infection and latency. For example, IE62 (the product of VZV gene 62) is a potent transactivator of many VZV genes associated with productive infection. While the function of IE63 (the product of VZV gene 63) is still uncertain, it may also function to regulate VZV gene expression and might be pivotal to the programmed switch from IE to early virus gene usage. During latency, however, the transactivation of VZV genes of IE62 or IE63 is modified. For example, in the presence of IE62 and IE63, the promoters for VZV genes 20, 21, 28, and 29 are active, while in latently infected ganglia where genes 62 and 63 are transcribed, the promoters for VZV genes 20 and 28 are silent. Thus, we will examine the hypothesis that IE62 and IE63 transactivation is regulated by VZV gene 21 and 29 proteins. Since the model relies on the translation of the transcripts detected during latency, we will search for proteins encoded by VZV genes 63, 62, 21 and 29 in the same latently infected human ganglionic cell. We will determine whether the VZV gene 21 protein forms a complex with VZV gene 29 protein, and whether this complex can inhibit VZV gene transactivation induced by IE62 or IE63. We will assay VZV replication, virus gene transcription, and virus promoter activity in cell lines expressing VZV genes 21 and 29. Our accumulated data on VZV gene expression in the human ganglia during latency have been assembled into a testable model designed to determine the interaction of these virus proteins. Such information will provide a rationale for future hypotheses about VZV reactivation, a cause of considerable morbidity, and occasional mortality, especially in elderly and immunocompromised humans.

潜伏性水痘带状疱疹病毒(VZV)背根再激活的研究 神经轴的所有水平的神经节产生严重的神经 疾病。在正常人潜伏感染的神经节中，我们已经证明 低水平的VZV DNA以环状形式存在，VZV基因 21、29、62和63被转录，VZV基因63被翻译。这个 事实上，在潜伏期转录的VZV基因包括那些立即- 已证实(基因62)或隐含(基因63)的早期(IE)基因 可能具有DNA结合能力的调节功能和基因(VZV 基因21和29)为我们的假设提供了理由 在潜伏期转录的VZV基因编码抑制病毒复制。 病毒蛋白的合成和感染性病毒粒子的组装是复杂的 依赖于协调基因活动的事件，并且在 生产性感染和潜伏期。例如，IE62(VZV的产品 基因62)是许多VZV基因的有效反式激活因子，这些基因与 生殖性感染。而IE63(VZV基因产物)的功能 63)仍不确定，它也可能具有调节VZV基因的功能 表达，并可能是从IE到早期编程转换的关键 病毒基因使用情况。然而，在潜伏期，VZV的反式激活 IE62或IE63的基因被修饰。例如，在IE62存在的情况下 和IE63是VZV基因20、21、28和29的启动子，而 在潜伏感染的神经节中，基因62和63被转录， VZV基因20和28的启动子是沉默的。因此，我们将研究 IE62和IE63反式激活受VZV基因21调控的假设 和29种蛋白质。由于该模型依赖于 在潜伏期检测到转录本，我们将搜索编码的蛋白质 VZV基因63、62、21和29在同一潜伏感染人群中的分布 神经节细胞。我们将确定VZV基因21蛋白是否形成 一种含有VZV基因29蛋白的复合体及其是否能抑制 IE62或IE63诱导的VZV基因反式激活。我们将对VZV进行检测 细胞内病毒复制、病毒基因转录和病毒启动子活性 表达VZV基因21和29的品系。我们积累的关于VZV基因的数据 潜伏期在人类神经节中的表达已经组装成一个 为确定这些病毒的相互作用而设计的可测试模型 蛋白质。这些信息将为未来的假设提供理论基础。 关于VZV的重新激活，这是一个相当大的发病率的原因，偶尔 死亡率，特别是老年人和免疫功能低下的人。