DARPin 与 HIV-1 CA 的特异性相互作用扰乱了膜中 Gag、RNA 包装和四跨膜超家族蛋白重塑的分布。

Specific Interaction of DARPin with HIV-1 CA Disturbs the Distribution of Gag, RNA Packaging, and Tetraspanin Remodelling in the Membrane.

机构信息

Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.

Department of Medical Technology, Division of Clinical Immunology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.

出版信息

Viruses. 2022 Apr 15;14(4):824. doi: 10.3390/v14040824.

A designed repeat scaffold protein (Ank1D4) recognizing the human immunodeficiency virus-1 (HIV-1) capsid (CA) was formerly established with antiviral assembly. Here, we investigated the molecular mechanism of Ank1D4 function during the late stages of the HIV-1 replication cycle. By applying stimulated emission-depletion (STED) microscopy, Gag polymerisation was interrupted at the plasma membrane. Disturbance of Gag polymerisation triggered Gag accumulation inside producer cells and trapping of the CD81 tetraspanin on the plasma membrane. Moreover, reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) experiments were performed to validate the packaging efficiency of RNAs. Our results advocated that Ank1D4 interfered with the Gag precursor protein from selecting HIV-1 and cellular RNAs for encapsidation into viral particles. These findings convey additional information on the antiviral activity of Ank1D4 at late stages of the HIV-1 life cycle, which is potential for an alternative anti-HIV molecule.

先前已经建立了一种设计的重复支架蛋白（Ank1D4），可识别人类免疫缺陷病毒-1（HIV-1）衣壳（CA），以进行抗病毒组装。在这里，我们研究了 Ank1D4 在 HIV-1 复制周期晚期的功能的分子机制。通过应用受激发射损耗（STED）显微镜，在质膜处中断 Gag 聚合。Gag 聚合的干扰会触发产生细胞内 Gag 的积累，并将 CD81 四跨膜蛋白固定在质膜上。此外，还进行了逆转录酶定量聚合酶链反应（RT-qPCR）实验以验证 RNA 的包装效率。我们的结果表明，Ank1D4 干扰了 Gag 前体蛋白，使其无法选择 HIV-1 和细胞 RNA 进行包装到病毒颗粒中。这些发现为 Ank1D4 在 HIV-1 生命周期晚期的抗病毒活性提供了更多信息，这为替代抗 HIV 分子提供了潜力。