SIV MOLECULAR CLONES W/ NEF MUTATIONS

SIV nef基因是维持SIV高表达的重要基因。 成年恒河猴的病毒载量与猴艾滋病进展（SAIDS） 猕猴 HIV和SIV的Nef基因序列高度保守 图案 这项研究是正在进行的功能分析的一部分， 体外Nef结构域（无细胞和组织培养测定系统） 和体内（用突变的SIV nef克隆感染的猕猴）。 目的SIV的nef基因对维持病毒的高表达具有重要意义 在成年恒河猴中的负荷和向猿类艾滋病（SAIDS）的进展。 HIV和SIV的Nef含有高度保守的src同源区3 （SH 3）-配体结构域，其特征在于pro-x-x-pro基序。 的 目的是确定该基序在病毒中的重要性， 致病性通过评估突变克隆，其中两个原残基 在猕猴体内突变为丙氨酸残基。 结果突变体 克隆SIVmac 239-P104 A/P107 A，在SH 3-配体中含有突变 结构域的Nef，是不能相互作用的Nef激活的激酶 (NAK)在体外激酶测定中。 猕猴被注射了这种疫苗 突变体，以测试该结构域的体内意义。 基因型和 在nef的表型逆转中观察到从 这些动物。 逆转与病毒的持久性相关， 发展到SAIDS。 因此，Nef与NAK的相互作用是 对病毒发病机制很重要。 未来的发展方向 与Nef的SH 3-配体结构域相互作用的蛋白质仍有待进一步研究。 鉴定 新的抗病毒策略可以基于 与Nef的SH 3配体结构域相互作用的药物。关键词SIV， 病毒致病

Significance The nef gene of SIV is important for maintaining high virus load and progression to simian AIDS (SAIDS) in adult rhesus macaques. Nef of HIV and SIV contains highly conserved sequence motifs. This sudy is part of an ongoing analysis of functional domains of Nef in vitro (cell-free and tissue culture assay systems) and in vivo (macaques infected with mutant SIV nef clones). Objectives The nef gene of SIV is important for maintaining high virus load and progression to simian AIDS (SAIDS) in adult rhesus macaques. Nef of HIV and SIV contains a highly conserved src-homology region 3 (SH3)-ligand domain characterized by the pro-x-x-pro motif. The objective was to determine the importance of this motif in viral pathogesis by evaluating a mutant clone, in which both pro residues were mutated to ala residues, in vivo in macaques. Results The mutant clone SIVmac239-P104A/P107A, containing mutations in the SH3-ligand domain of Nef, was not able to interact with the Nef-activated kinase (NAK) in in vitro kinase assays. Macaques were inoculated with this mutant to test the in vivo significance of this domain. Genotypic and phenotypic reversions in nef were observed in viruses recovered from these animals. The reversions correlated with viral persistence and progression to SAIDS. Thus, the interaction of Nef with NAK is important for viral pathogenesis. Future Directions The cellular protein(s) interacting with the SH3-ligand domain of Nef remains to be identified. Novel antiviral strategies can be developed based on drugs that interact with the SH3-ligand domain of Nef. KEYWORDS SIV, viral pathogenesis