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目的研究HIV-1整合酶与细胞蛋白Importinα1(Impα1)的相互作用及其对HIV-1复制的影响。方法采用化学发光免疫共沉淀系统定量检测细胞内HIV-1整合酶与Impα1蛋白的相互作用,同时利用基因沉默技术下调CD4+C8166 T细胞中Impα1的表达,以研究Impα/β入核转运途径对HIV-1复制的影响。结果当T细胞内Impα1表达下降时,HIV-1对其感染性显著减弱。Real-time PCR分析结果显示,尽管病毒逆转录未受到影响,但其2-LTR DNA(入核指标)及整合DNA的形成受到了抑制,提示Impα1的下调明显影响HIV-1入核转运过程。结论该研究为HIV-1整合酶与Impα1蛋白之间的相互作用及其在病毒感染中的影响提供了重要的实验依据。
Objective To investigate the interaction between HIV-1 integrase and the cellular protein Importinα1 (Impα1) and its effect on HIV-1 replication. Methods The chemiluminescent co-immunoprecipitation system was used to quantitatively detect the interaction between HIV-1 integrase and Impα1 protein in the cells. At the same time, the gene silencing technique was used to down-regulate the expression of Impα1 in CD4 + C8166 T cells to investigate the effects of Impα / The impact of HIV-1 replication. Results When the expression of Impα1 in T cells decreased, the infectivity of HIV-1 was significantly weakened. Real-time PCR analysis showed that although the reverse transcription of the virus was not affected, the formation of 2-LTR DNA (nuclear import index) and DNA integration were inhibited, suggesting that downregulation of Impα1 significantly affected the translocation of HIV-1 into the nucleus. Conclusion This study provides an important experimental evidence for the interaction between HIV-1 integrase and Impα1 protein and its effect on viral infection.