The regulator of expression of virion (Rev) protein binds specifically to the Rev-responsive element (RRE) RNA in order to regulate the expression of the human immunodeficiency virus (HIV)-1 genes. Fluorescence indicator displacement assays have been used to identify ligands that can inhibit the Rev–RRE interaction; however, the small fluorescence indicators cannot fully replace the Rev peptide or protein. As a result, a single rhodamine B labeled Rev (RB-Rev) model peptide was utilized in this study to develop a direct and efficient Rev–RRE inhibitor screening model. Due to photon-induced electron transfer quenching of the tryptophan residue on the RB fluorophore, the fluorescence of RB in Rev was weakened and could be dramatically reactivated by interaction with RRE RNA in ammonium acetate buffer (approximately six times). The interaction could reduce the electron transfer between tryptophan and RB, and RRE could also increase RB fluorescence. The inhibitor screening model was evaluated using three known positive Rev–RRE inhibitors, namely, proflavin, 6-chloro-9-[3-(2-chloroethylamino)propylamino]-2-methoxyacridine (ICR 191), and neomycin, as well as a negative drug, arginine. With the addition of the positive drugs, the fluorescence of the Rev–RRE decreased, indicating the displacement of RB-Rev. This was confirmed using atomic force microscopy (AFM) and the fluorescence was essentially unaffected by the addition of arginine. The results demonstrated that RB-Rev can be used as a fluorescent probe for recognizing small ligands that target RRE RNA. The Rev–RRE inhibitor screening model offers a novel approach to evaluating and identifying long-acting Rev inhibitors.

病毒颗粒 (Rev) 蛋白表达调节剂与 Rev 反应元件 (RRE) RNA 特异性结合，以调节人类免疫缺陷病毒 (HIV)-1 基因的表达。荧光指示剂置换分析已被用于鉴定可抑制 Rev-RRE 相互作用的配体；然而，小荧光指示剂并不能完全替代 Rev 肽或蛋白质。因此，本研究利用单一罗丹明 B 标记的 Rev (RB-Rev) 模型肽来开发直接有效的 Rev-RRE 抑制剂筛选模型。由于 RB 荧光团上色氨酸残基的光子诱导电子转移猝灭，Rev 中 RB 的荧光减弱，并且可以通过与醋酸铵缓冲液中的 RRE RNA 相互作用（大约六次）显着重新激活。这种相互作用可以减少色氨酸和RB之间的电子转移，RRE也可以增加RB的荧光。使用三种已知的阳性 Rev-RRE 抑制剂评估抑制剂筛选模型，即原黄素、6-氯-9-[3-(2-氯乙氨基)丙氨基]-2-甲氧基吖啶 (ICR 191) 和新霉素，以及一种阴性药物，精氨酸。随着阳性药物的加入，Rev-RRE 的荧光减少，表明 RB-Rev 发生了位移。使用原子力显微镜 (AFM) 证实了这一点，荧光基本上不受添加精氨酸的影响。结果表明，RB-Rev 可用作识别靶向 RRE RNA 的小配体的荧光探针。Rev-RRE 抑制剂筛选模型提供了一种评估和鉴定长效 Rev 抑制剂的新方法。