STRUCTURAL AND FUNCTIONAL STUDIES OF MAMMALIAN ENDOGENOUS RETROVIRAL SEQUENCES

哺乳动物内源逆转录病毒序列的结构和功能研究

• 批准号: 3790737
• 负责人: A S KHAN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3790737
• 关键词: Retroviridae; enzyme mechanism; genetic manipulation; genetic strain; murine leukemia virus; nucleic acid sequence; transposon /insertion element; viral leukemogenesis; virus DNA; virus genetics; virus infection mechanism

The integrase genes of leukemogenic MCF13 and non-leukemogenic MCFM111A murine retroviruses were compared in their structure and biological activities. Nucleotide sequence analysis indicated that the two integrase genes were distinct: there were 21 substituted amino acids between MCF13 and MCFM111A, of which 4 affected the hydropathicity profile; in addition, 4 amino acids were present in MCF13 murine leukemia virus (MuLV) which were absent in MCFM111A MuLV. To determine whether the structural differences seen between the integrase genes of leukemogenic versus non-leukemogenic murine viruses affected integrase function, we analyzed the integrase protein activities using an in vitro biological system. This assay measured the incorporation of linear viral DNAs, present in cytoplasmic extracts from infected cells, into phi-X174 target DNA. The results indicated that integration by leukemogenic MCF13 was two-fold greater than by non-leukemogenic MCFM111A.

致白血病MCF13和非致白血病MCFM111A的整合酶基因 比较了小鼠逆转录病毒的结构和生物学特性 活动。 核苷酸序列分析表明，两者 整合酶基因不同：有 21 个取代氨基酸 MCF13和MCFM111A之间，其中4个影响亲水性 轮廓;此外，MCF13 鼠中存在 4 种氨基酸 MCFM111A MuLV 中不存在白血病病毒 (MuLV)。 确定 整合酶基因之间是否存在结构差异 致白血病与非致白血病鼠病毒影响整合酶 功能，我们使用体外分析了整合酶蛋白的活性 生物系统。 该测定测量了线性的掺入 病毒 DNA 存在于受感染细胞的细胞质提取物中， phi-X174 目标 DNA。 结果表明，积分通过 致白血病 MCF13 是非致白血病的两倍 MCFM111A。