APOBEC3G/CEM15 Inhibition of Lentivirus Replication

APOBEC3G/CEM15 抑制慢病毒复制

基本信息

批准号：
8679134
负责人：
Nathaniel R. Landau
金额：
$ 26.7万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-16 至 2015-08-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8679134
关键词：
Address Antibodies Antiviral Agents Binding Binding Sites Biology CD4 Positive T Lymphocytes Cells Cytidine Deaminase Cytosine Nucleotides Deaminase Defect Endogenous Retroviruses Evolution Family Family member Gene Expression Gene Family Genes Genetic Transcription HIV HIV-1 Human Human Genome Immune response Immune system Individual Knock-out Knockout Mice Learning Light Mediating Molecular Profiling Monoclonal Antibodies Mus Pattern Physiological Physiology Plasmids Play Protein Family Proteins Relative (related person)Reporter Retroelements Retrotransposon Retroviridae Role SIV Specificity Subfamily lentivirinae T-Lymphocyte Testing Tissues Transcript Transcriptional Regulation Transgenic Mice Viral Virion Virus Virus Replication cell type cytokine in vivo insight knock-down macrophage mouse model mutant promoter stable cell line transcription factor

项目摘要

The 7 APOBEC3 genes (APOBEC3A, B, C, DE, F, G and H) in the human genome encode antiviral cytidine deaminases. The deaminases are specifically packaged into virions where they deaminate cytosine nucleotides in the minus-strand of the viral reverse transcripts. The Vif accessory protein counteracts APOBEC3F and G by binding and inducing their degradation. The project will investigate several features APOBEC3 biology. Specific aim 1 will generate a panel of monoclonal antibodies against the family members and use these to characterize the tissue and cell-type expression of the individual family members. Each family member will be tested in stable cell lines to evaluate their antiviral activity against HIV-1 and SIV. Specific aim 2 will investigate the transcriptional regulation of the APOBEC3 genes. Microarray expression profiling will be used to identify the transcription factors that regulate transcription of the APOBEC3 genes by finding expression differences between closely matched nonpermissive and permissive cells. Cellular genes will be identified that are co-regulated with APOBEC3G. Transcription factor binding sites in the APOBEC3G promoter will be identified to define how the gene is regulated and to provide insight into ways to influence promoter activity that would increase the intracellular levels of protective APOBEC3 proteins. Specific aim 3 will investigate the mechanisms by which APOBEC3 proteins are packaged into virions will study the role of APOBEC3 in the mouse which has only a single APOBEC3 gene. Specific aim 4 will use the mouse model to study the role of APOBEC3 in the immune response. The studies will make use of an APOBEC3 knock-out mouse to investigate the mechanism by which APOBEC3 suppresses retrovirus replication in vivo and whether the protein plays a role in the suppression of retrotransposons or a yet unidentified role in the immune system.

人基因组中的7个APOBEC3基因（Apobec3a，b，d，f，g和h）编码抗病毒胞苷脱氨酶。脱氨酶被专门包装到病毒体中，它们在病毒逆转录物的负链中脱氨基核苷酸。 VIF辅助蛋白通过结合和诱导其降解来抵消APOBEC3F和G。该项目将研究APOBEC3生物学的几种功能。具体的目标1将生成针对家庭成员的一组单克隆抗体，并使用这些抗体来表征单个家庭成员的组织和细胞类型表达。每个家庭成员将在稳定的细胞系中进行测试，以评估其针对HIV-1和SIV的抗病毒活性。具体目标2将研究APOBEC3基因的转录调控。微阵列表达分析将用于确定通过在密切匹配的非耐药和允许细胞之间找到表达差异来调节APOBEC3基因转录的转录因子。将鉴定出与Apobec3g共同调节的细胞基因。将确定APOBEC3G启动子中的转录因子结合位点，以定义基因的调节方式，并提供对影响启动子活性的方法，从而增加保护性APOBEC3蛋白的细胞内水平。具体目标3将研究将APOBEC3蛋白包装到病毒体中的机制，将研究Apobec3在仅具有单个APOBEC3基因的小鼠中的作用。特定的目标4将使用小鼠模型研究APOBEC3在免疫反应中的作用。该研究将利用APOBEC3敲除小鼠研究APOBEC3抑制体内逆转录病毒复制的机制，以及该蛋白质是否在抑制逆转转座子的抑制或在免疫系统中尚未识别的作用中起作用。