Repair of HCMV-Induced DNA Damage in Infected Cells

修复受感染细胞中 HCMV 诱导的 DNA 损伤

• 批准号: 7155531
• 负责人: ELIZABETH A FORTUNATO
• 金额: $ 24.45万
• 依托单位: UNIVERSITY OF IDAHO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-15 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7155531
• 关键词: Affect; Antibodies; Apoptosis; Biological Assay; Birth; Blindness; Cell Cycle; Cell Cycle Proteins; Cells; Chromosomal Breaks; Chromosomes; Chromosomes, Human, Pair 1; Comet Assay; Complex; Congenital Abnormality; Coupled; Cytogenetics; Cytomegalovirus; DNA; DNA Damage; DNA Repair; DNA Sequence; Data; Development; Environment; Fibroblasts; Fluorescent in Situ Hybridization; Genome; Genus Capra; Goat; Head; Healed; In Vitro; Infant; Infection; Learning Disabilities; Life; Literature; Localized; Mental Retardation; Methods; Microcephaly; Mitotic; Monitor; Morbidity - disease rate; Nervous System Trauma; Neuraxis; Neurons; Newborn Infant; Nonhomologous DNA End Joining; Nuclear; Phase; Population; Positioning Attribute; Protein p53; Proteins; Pulsed-Field Gel Electrophoresis; Tail; Techniques; Testing; Thinking; Time; Ultraviolet Rays; Viral; Viral Antigens; Viral Proteins; Virus; Work; cell motility; cell type; clinically relevant; congenital infection; deafness; dimer; healing; hearing impairment; human H2AX protein; mortality; protein expression; relating to nervous system; repaired; response; tissue culture

Human Cytomegalovirus (HCMV) is the major viral cause of birth defects, infecting 1-2% of all newborns annually. Approximately 5-10% of these congenitally infected infants will manifest signs of serious neurological damage at birth, which can include deafness, blindness, mental retardation and microcephaly. Another 10-15% will develop sensori-neural hearing loss and/or learning disabilities within the first 10 years of life. Our long-term goat is to understand the mechanism behind the development of morbidity and mortality in infants congenitally infected with HCMV. Over the last several years we have studied the interaction of HCMV with the cell cycle and DNA repair machinery of the permissively infected cell. We have discerned that HCMV sequesters many key regulatory and repair proteins into its viral replication centers. However, it appears to partition the components of several complexes so that all the proteins are present within the replication centers, but not all are available to the cellular genome. We have also determined that HCMV can induce specific damage on chromosome 1 during S-phase. When coupled with the literature regarding nonspecific damage induced at late times post infection our data more clearly highlights the genotoxic effects of HCMV. We hypothesize that long-term detrimental consequences to the cellular genome may occur if 1) the initial specific damage is propagated or 2) damage incurred at late times post infection is not repaired due to sequestration of the repair machinery. To test our hypothesis, we propose three specific aims. First, we will thoroughly define the parameters of chromosome lq breakage in HCMV-infected cells with regard to rapidity of induction and cell cycle phase at time of infection. We think it is imperative that our results be moved into more clinically relevant cell types, especially cells of neural lineage, as these are the cells most severely affected by the virus during congenital infection. Second, we also will determine the consequences of chromosome 1 damage in these clinically relevant cells, and whether in a semi-permissive environment we can observe propagation of the chromosome 1 damage instead of healing of the break or movement of the cell toward apoptosis. Lastly, we will characterize the ability of HCMV-infected cells to repair exogenously introduced damage at late times post infection, after viral replication centers are assembled.

人类巨细胞病毒(HCMV)是导致出生缺陷的主要病毒原因，感染了1-2%的新生儿 每年一次。这些先天感染的婴儿中大约有5%-10%会表现出严重的 出生时的神经损伤，可能包括耳聋、失明、智力低下和小头畸形。 另有10%-15%的人将在头10年内发展为感觉神经性听力损失和/或学习障碍 生活的一部分。我们的长远目标是了解发病和发病背后的机制。 先天性感染巨细胞病毒的婴儿死亡率。在过去的几年里，我们研究了 巨细胞病毒与允许感染细胞的细胞周期和DNA修复机制的相互作用。我们有 发现HCMV将许多关键的调节和修复蛋白隔离到其病毒复制中心。 然而，它似乎分割了几个复合体的成分，使所有的蛋白质都存在 在复制中心内，但不是所有的都可用于细胞基因组。我们还确定， 人巨细胞病毒可在S期引起1号染色体的特异性损伤。 当结合关于感染后晚期引起的非特异性损伤的文献时，我们的 数据更清楚地突出了人巨细胞病毒的遗传毒性效应。我们假设，从长远来看， 如果1)最初的特定损伤是传播的或2)损伤，则可能发生对细胞基因组的后果 感染后后期发生的感染由于修复机械的隔离而无法修复。测试我们的 假设，我们提出了三个具体目标。首先，我们将彻底定义染色体的参数 人巨细胞病毒感染细胞LQ断裂与诱导速度和细胞周期时相的关系 感染。我们认为，我们的结果必须转移到更多临床相关的细胞类型中，特别是 神经系细胞，因为这些细胞是先天感染期间受病毒影响最严重的细胞。 其次，我们还将确定1号染色体损伤在这些临床相关疾病中的后果 细胞，以及在半允许的环境中，我们是否可以观察到1号染色体的传播 损伤而不是愈合细胞的断裂或走向凋亡的运动。最后，我们将 人巨细胞病毒感染细胞在后期修复外源性损伤的能力 感染，在病毒复制中心组装后。