HIV-1整合酶与宿主动力蛋白中间链1蛋白间相互作用的研究

摘要

目的 研究在1型人类免疫缺陷病毒(HIV-1)的复制过程中,HIV-1整合酶(HIV-1 IN)与细胞质动力蛋白复合体中的动力蛋白中间链1蛋白(DIC1)之间的相互作用.方法 本实验主要采用转染,免疫共沉淀(Co-Immunoprecipitation,Co-IP),免疫荧光(Immunofluorescence)和蛋白质印记法(Western Blot)对HIV-1 IN和DIC1进行检测.结果 通过免疫共沉淀法显示HIV-1 IN与DIC1的相互作用,以及HIV-1 IN C末端的重要作用.通过免疫荧光法,证实DIC1和IN是在细胞质中相结合的,而非在细胞核中结合.结论 HIV-1进入细胞后可以立即与动力蛋白复合体相结合,从而促进病毒向细胞核移动以及病毒DNA整合.%Objective To investigate the corelation of HIV-1 integrase (IN) with the dynein intermediate chain 1 (DIC1) during the replication cycle of Human Immunodeficiency Virus type 1 (HIV-1).Methods HIV-1 IN and DIC1 were checked by transfection,Co-Immunoprecipitation (Co-IP),immunofluorescence and western blotting.Results Co-immunoprecipitation approach was performed to reveal that HIV-1 IN was associated with another protein,DIClof the cytoplasmic dynein motor complex,and the C-terminus of IN was essential for this interaction.The immunofluorescence assay also showed that DIC1 was co-localized with HIV IN in the cytoplasm,but not in the nucleus,suggesting that HIV-1 IN was associated with dynein motor complex in the cytoplasm.Conclusion This study provides evidence that HIV enters cells and correlates with the dynein motor complex through viral protein IN and DIC1 interaction,which facilitates its movement to the nucleus for viral DNA integration.