NOVEL CELL CYCLE REGULATION BY ADENOVIRUS E1B 55K

腺病毒 E1B 55K 对细胞周期的新型调控

• 批准号: 6464597
• 负责人: David Arnold Ornelles
• 金额: $ 3.01万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6464597
• 关键词: Adenoviridae; cell cycle; flow cytometry; gene mutation; genetic transcription; intracellular transport; messenger RNA; oncoproteins; protein structure function; tissue /cell culture; transmission electron microscopy; viral carcinogenesis; virus RNA; virus infection mechanism; virus protein

The adenovirus E1B 55-kDa (55K) protein functions during virus growth and virus-mediated cellular transformation. During virus growth, the 55K protein binds the E4orf6 protein and regulates mRNA biogenesis. During transformation, the 55K protein binds the tumor suppressor p53 and blocks transcription from p53-responsive genes including those that promote apoptosis and induce G1 growth arrest. However, most cells infected with the 55K-mutant virus do not undergo apoptosis, do not arrest in G1, and do not fail to replicate mutant viral DNA. Consequently, the link between 55K function in virus growth and transformation remains unclear. This work will elucidate this link as part of the larger goal of understanding the means by which adenovirus targets mechanisms of cellular growth control for virus growth. Because 55K-mutant viruses grow only in HeLa cells that were infected in early S phase but not in G1, the 55K protein overcomes a restriction imposed on virus growth by the cell cycle. This restriction and the 55K function that overcomes this restriction will be determined by three specific aims. First, the hypothesis that adenovirus must usurp the control of mRNA for cell cycle-independent growth will be evaluated by comparing the growth of related G1-restricted adenovirus mutants and the control of mRNA transport among infected cells that are permissive and restrictive for virus growth. Second, heterokaryons of permissive and restrictive cells will be used to elucidate the cellular basis for this restriction and determine the dominance of the restrictive phenotype. Third, cellular proteins targeted by 55K/E4orf6 complex for cell cycle-independent virus growth will be identified and the cell cycle- regulation of these proteins determined. This study will determine how the 55K protein subverts cell cycle controls for virus growth. This may represent the first link between the roll of the 55K protein in lytic growth and viral transformation. These studies will increase our understanding of the mechanisms of viral pathogenesis and viral oncogenesis as well as fundamental mechanisms of cellular growth control. Furthermore, this study will identify adenovirus mutants that are restricted for growth in S phase cells. Such replication competent viruses can be used as oncolytic agents to treat rapidly growing human tumors.

腺病毒 E1B 55-kDa (55K) 蛋白在病毒生长和病毒介导的细胞转化过程中发挥作用。 在病毒生长过程中，55K 蛋白结合 E4orf6 蛋白并调节 mRNA 生物发生。在转化过程中，55K 蛋白结合肿瘤抑制因子 p53，并阻断 p53 反应基因的转录，包括促进细胞凋亡和诱导 G1 生长停滞的基因。 然而，大多数感染 55K 突变病毒的细胞不会发生凋亡，不会停滞在 G1 期，并且不会无法复制突变病毒 DNA。因此，55K 在病毒生长和转化中的功能之间的联系仍不清楚。 这项工作将阐明这种联系，作为了解腺病毒针对病毒生长的细胞生长控制机制的更大目标的一部分。由于 55K 突变病毒仅在早期 S 期感染的 HeLa 细胞中生长，而不在 G1 期感染，因此 55K 蛋白克服了细胞周期对病毒生长的限制。 这一限制以及克服这一限制的 55K 功能将由三个具体目标决定。 首先，通过比较相关 G1 限制性腺病毒突变体的生长和允许和限制病毒生长的受感染细胞之间 mRNA 运输的控制，将评估腺病毒必须篡夺细胞周期独立生长的 mRNA 控制的假设。 其次，允许性和限制性细胞的异核体将用于阐明这种限制的细胞基础并确定限制性表型的优势。第三，将鉴定55K/E4orf6复合物针对细胞周期非依赖性病毒生长所靶向的细胞蛋白，并确定这些蛋白的细胞周期调节。这项研究将确定 55K 蛋白如何破坏病毒生长的细胞周期控制。 这可能代表了 55K 蛋白在裂解生长和病毒转化中的滚动之间的第一个联系。这些研究将增加我们对病毒发病机制和病毒肿瘤发生机制以及细胞生长控制基本机制的理解。此外，这项研究将鉴定出限制在 S 期细胞中生长的腺病毒突变体。 这种具有复制能力的病毒可用作溶瘤剂来治疗快速生长的人类肿瘤。