Abstract：
To integrate with host chromatin and establish a productive infection, HIV-1 must translocate the viral Ribonucleoprotein (RNP) complex through the nuclear pore complex (NPC). Current assay to measure HIV-1 nuclear import relies on a transient byproduct of HIV-1 integration failure called 2-LTR circles. However, 2-LTR circles require complete or near-complete reverse transcription and association with the non-homologous end joining (NHEJ) machinery in the nucleus, which can complicate interpretation of 2-LTR circle formation as a measure of nuclear import kinetics. Here, we describe an approach to measure nuclear import of infectious HIV-1 particles. This involves chemically induced dimerization of Nup62, a central FG containing nucleoporin. Using this technique, nuclear import of infectious particles can be monitored in both primary and cell culture models. In response to host factor depletion or restriction factors, changes in HIV-1 nuclear import can be effectively measured using the nuclear import kinetics (NIK) assay.
摘要：
与宿主染色质整合并建立生产性感染，HIV-1必须通过核孔复合物（NPC）转运病毒核糖核蛋白（RNP）复合物。当前用于测量HIV-1核输入的测定依赖于称为2-LTR环的HIV-1整合失败的瞬时副产物。然而,2-LTR环需要完全或接近完全的逆转录，并与细胞核中的非同源末端连接（NHEJ）机制相关联，这可能会使2-LTR环形成的解释复杂化，以衡量核输入动力学。这里,我们描述了一种方法来测量传染性HIV-1颗粒的核输入。这涉及化学诱导的Nup62二聚化，Nup62是一种含有核孔蛋白的中央FG。使用这种技术，可以在原代和细胞培养模型中监测感染性颗粒的核输入。响应宿主因子消耗或限制因子，使用核输入动力学（NIK）测定法可以有效地测量HIV-1核输入的变化。
