APOBEC3G/CEM15 Inhibition of Lentivirus Replication

描述（由申请人提供）：人类基因组中的7个APOBEC 3基因（APOBEC 3A、B、C、DE、F、G和H）编码抗病毒胞苷脱氨酶。脱氨酶被特异性地包装到病毒体中，在病毒体中它们使病毒反向转录物的负链中的胞嘧啶核苷酸脱氨。Vif辅助蛋白通过结合并诱导其降解来抵消APOBEC 3F和G。该项目将研究APOBEC 3生物学的几个特征。具体目标1将产生一组针对家族成员的单克隆抗体，并使用这些抗体来表征单个家族成员的组织和细胞类型表达。每个家族成员将在稳定的细胞系中进行测试，以评估其对HIV-1和SIV的抗病毒活性。具体目标2将研究APOBEC 3基因的转录调控。微阵列表达谱分析将用于通过发现紧密匹配的非允许细胞和允许细胞之间的表达差异来鉴定调节APOBEC 3基因转录的转录因子。将鉴定与APOBEC 3G共调节的细胞基因。将鉴定APOBEC 3G启动子中的转录因子结合位点，以确定基因是如何调控的，并提供对影响启动子活性的方法的了解，从而增加保护性APOBEC 3蛋白的细胞内水平。具体目标3将研究APOBEC 3蛋白被包装到病毒体中的机制，将研究APOBEC 3在只有单个APOBEC 3基因的小鼠中的作用。具体目标4将使用小鼠模型来研究APOBEC 3在免疫应答中的作用。这些研究将利用APOBEC 3基因敲除小鼠来研究APOBEC 3在体内抑制逆转录病毒复制的机制，以及该蛋白质是否在抑制逆转录转座子中发挥作用或在免疫系统中发挥尚未确定的作用。 APOBEC 3蛋白是一个由7种蛋白组成的家族，限制包括HIV在内的病毒的复制。该项目将研究APOBEC 3家族成员的作用，以了解哪些成员对HIV-1具有活性，以及它们如何干扰病毒的复制。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The APOBEC3 proteins are a family of 7 proteins that restrict the replication of viruses including HIV. The project will investigate the roles of the APOBEC3 family members to understand which ones are active against HIV-1 and how they interfere with the replication of the virus.