APOBEC3G/CEM15 Inhibition of Lentivirus Replication

APOBEC3G/CEM15 抑制慢病毒复制

• 批准号: 7926684
• 负责人: Nathaniel R. Landau
• 金额: $ 7.04万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-26 至 2010-12-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7926684
• 关键词: Binding Sites; Biological Assay; Biology; Cell Line; Cells; Cercopithecus pygerythrus; Chimera organism; Cloning; Complementary DNA; Cytidine Deaminase; DNA; Defect; Development; Enzymes; Family; Goals; HIV; HIV-1; Homologous Gene; Hospitals; Human; Immune system; In Vitro; Infection; Knockout Mice; London; Lymphocyte; Lymphocyte Subset; Mediating; Messenger RNA; Murine leukemia virus; Mus; Mutate; Pharmaceutical Preparations; Phenotype; Physiological; Primates; Proteins; Proviruses; RNA; RNA Binding; RNA Editing; Rodent; Role; Site; Specificity; Subfamily lentivirinae; System; T-Lymphocyte; Testing; Therapeutic; Transcript; Viral; Virion; Virus; Virus Replication; cofactor; immune function; inhibitor/antagonist; insight; interest; member; novel; small molecule; therapeutic development; viral RNA

DESCRIPTION (provided by applicant): The lentivirus virion infectivity factor (Vif) is an accessory protein that is required for productive replication of the virus in primary cells and some, but not all, transformed T cell lines. HIV-1 that is genetically deficient in Vif (delta-Vif) fails to replicate in nonpermissive cells as the result of the expression of the cell protein APOBEC3G/CEM15. APOBEC3G is a member of the cytidine deaminase family of RNA editing enzymes. Simpler viruses such as murine leukemia virus do not encode Vif. Nevertheless, cloning and functional analysis showed that mouse APOBEC3G is a potent inhibitor of delta-Vif and wild-type HIV-1. Similarly, African Green monkey APOBEC3G was also active against wild-type and delta-Vif HIV-1 but only inhibited delta-Vif but not SIVagm. These findings suggest a species-specific interaction between Vif and APOBEC3G. The goals of this project are to understand the mechanism by which APOBEC3G reduces the infectivity of delta-Vif HIV-1 and to understand how Vif alleviates this inhibition. Human:AGM chimeric APOBEC3Gs will be constructed and used to determine the domain that determines the interaction. HIV-1 will be adapted to replicate in cells expressing non-homologous APOBEC3G. Physical interaction of APOBEC3G and Vif with each other and with cellular factors will be detected by coimmunoprecipitation. To identify a viral editing substrate for APOBEC3G, encapsidated viral RNA, viral mRNA, viral cDNA and proviral DNA will be sequenced from wild-type and delta-Vif virus grown in the presence or absence of APOBEC3G. In addition, to understand the role physiological role of APOBEC3G, knock-out mice will be constructed. Several immune function assays will be used to determine the effects of the deficiency on the immune system. Insights provided by these studies will have implications regarding targeting the APOBEC3G:Vif interaction for the development of novel HIV therapeutics. Such drugs would interfere with the APOBEC3G:Vif interaction without inhibiting APOBEC3G function.

描述（由申请人提供）：慢病毒病毒感染因子（VIF）是一种辅助蛋白，是对原代细胞中病毒的生产性复制所必需的，有些（但不是全部）转化为T细胞系。由于细胞蛋白APOBEC3G/CEM15的表达，VIF（Delta-VIF）在遗传上缺陷（Delta-VIF）的HIV-1未能复制。 APOBEC3G是RNA编辑酶的胞苷脱氨酶家族的成员。诸如鼠白血病病毒之类的简单病毒不编码VIF。然而，克隆和功能分析表明，小鼠APOBEC3G是Delta-VIF和野生型HIV-1的有效抑制剂。同样，非洲绿色猴子Apobec3g也对野生型和Delta-VIF HIV-1也很活跃，但仅抑制Delta-VIF，但不抑制Sivagm。这些发现表明VIF和APOBEC3G之间的物种特异性相互作用。该项目的目标是了解APOBEC3G降低Delta-VIF HIV-1的感染性的机制，并了解VIF如何减轻这种抑制作用。人：将构建AGM嵌合apobec3gs并用于确定确定相互作用的域。 HIV-1将适用于表达非同源apobec3g的细胞中复制。 APOBEC3G和VIF与彼此以及与细胞因子的物理相互作用将通过共免疫沉淀检测。为了鉴定apobec3g的病毒编辑底物，将在存在或不存在Apobec3g的情况下生长的野生型和Delta-VIF病毒对封装的病毒RNA，病毒mRNA，病毒cDNA和前病毒DNA进行测序。此外，要了解Apobec3g的生理作用，将构建敲除小鼠。几种免疫功能分析将用于确定缺乏对免疫系统的影响。这些研究提供的见解将对靶向apobec3g：VIF的相互作用具有含义。此类药物会干扰Apobec3g：VIF相互作用而不会抑制APOBEC3G功能。