High-affinity interaction between HIV-1 Vpr and specific sequences that span the C/EBP and adjacent NF-κB sites within the HIV-1 LTR correlate with HIV-1-associated dementia
High-affinity interaction between HIV-1 Vpr and specific sequences that span the C/EBP and adjacent NF-κB sites within the HIV-1 LTR correlate with HIV-1-associated dementia
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DOI：
10.1089/104454904773819842
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发表时间：
2004-04-01
影响因子：
3.1
通讯作者：
Wigdahl, B
中科院分区：
文献类型：
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作者：
Burdo, TH;Nonnemacher, M;Wigdahl, B
许多宿主和病毒因素可能参与了HIV-1相关性痴呆（HIVD）的发病和进展。先前的研究表明，在单核细胞/巨噬细胞谱系的驻留中枢神经系统（CNS）细胞中的病毒基因表达在神经毒性病毒蛋白和感染性病毒的产生、细胞基因表达的失调和/或神经胶质细胞和神经元细胞群的功能障碍中起核心作用。HIV-1复制部分地通过细胞转录因子与病毒反式激活因子达特和病毒蛋白R（Vpr）之间的相互作用以及LTR内的顺式作用启动子元件来调节。我们以前已经证明，Vpr结合CCAAT/增强子结合蛋白（C/EBP）站点I和紧邻该站点的下游序列内的选定序列配置具有高亲和力。本文报告的研究确立了HIVD诊断与HIV-1 LTR流行率增加之间的相关性，该LTR含有C/EBP结合位点I，对Vpr表现出高亲和力。为此，Vpr与C/EBP位点I变异体在47个LTR从3个非痴呆患者和96个LTR从7个痴呆患者的相互作用进行了检查。利用竞争电泳迁移率变动（EMS）分析来检查Vpr结合到含有C/EBP位点I变体的寡核苷酸探针。我们证明，89%的LTRs来自临床痴呆患者的C/EBP网站I配置，表现出较高的相对亲和力Vpr，而只有11%的LTRs包含C/EBP网站I配置，表现出较低的相对亲和力Vpr结合表型。相比之下，对缺乏临床明显痴呆的患者的LTR的检查显示，只有53%的脑源性LTR含有C/EBP位点I构型，其对Vpr显示出高相对亲和力，而47%的LTR含有C/EBP位点I构型，其显示出低相对亲和力Vpr结合表型。我们建议，顺式作用元件之间的序列特异性相互作用的LTR，C/EBP家族成员的转录因子，和病毒体相关的反式激活蛋白Vpr在HIVD的发病机制中发挥重要作用。
Numerous host and viral factors likely participate in the onset and progression of HIV-1-associated dementia (HIVD). Previous studies have suggested that viral gene expression in resident central nervous system (CNS) cells of monocyte/macrophage lineage play a central role in the production of neurotoxic viral proteins and infectious virus, deregulation of cellular gene expression, and/or dysfunction of glial and neuronal cell populations. HIV-1 replication is regulated, in part, by interactions between cellular transcription factors and the viral trans-activators, Tat and viral protein R (Vpr), with cis-acting promoter elements within the LTR. We have previously demonstrated that Vpr binds with high affinity to selected sequence configurations within CCAAT/enhancer binding protein (C/EBP) site I and downstream sequences immediately adjacent to this site. Studies reported herein establish a correlation between the diagnosis of HIVD and the increased prevalence of HIV-1 LTRs containing a C/EBP binding site, I that exhibits high affinity for Vpr. To this end, the interaction of Vpr with C/EBP site I variants in 47 LTRs from three nondemented patients and 96 LTRs from seven demented patients was examined. Competition electrophoretic mobility shift (EMS) analyses were utilized to examine Vpr binding to oligonucleotide probes containing C/EBP site I variants. We demonstrated that 89% of LTRs derived from patients exhibiting clinical dementia contained C/EBP site I configurations that displayed a high relative affinity for Vpr, while only 11% of LTRs contained C/EBP site I configurations that exhibited a low relative affinity Vpr binding phenotype. In contrast, examination of LTRs derived from patients lacking clinically evident dementia revealed that only 53% of brain-derived LTRs contained C/EBP site I configurations that displayed a high relative affinity for Vpr, while 47% of LTRs contained C/EBP site I configurations that exhibited a low relative affinity Vpr binding phenotype. We propose that sequence-specific interactions between cis-acting elements in the LTR, members of the C/EBP family of transcription factors, and the virion-associated trans-activator protein Vpr play important roles in the pathogenesis of HIVD.