衣壳
核运输
核孔
细胞核
细胞生物学
生物
抄写(语言学)
核心
细胞质
转录因子
核定位序列
逆转录酶
病毒复制
核糖核酸
病毒学
病毒
基因
遗传学
哲学
语言学
作者
Adarsh Dharan,Niklas Bachmann,Sarah Talley,Virginia Zwikelmaier,Edward M. Campbell
出处
期刊:Nature microbiology
日期:2020-06-01
卷期号:5 (9): 1088-1095
被引量:152
标识
DOI:10.1038/s41564-020-0735-8
摘要
Retroviral infection involves the reverse transcription of the viral RNA genome into DNA, which is subsequently integrated into the host cell genome. Human immunodeficiency virus type 1 (HIV-1) and other lentiviruses mediate the infection of non-dividing cells through the ability of the capsid protein1 to engage the cellular nuclear import pathways of the target cell and mediate their nuclear translocation through components of the nuclear pore complex2–4. Although recent studies have observed the presence of the capsid protein in the nucleus during infection5–8, reverse transcription and disassembly of the viral core have conventionally been considered to be cytoplasmic events. Here, we use an inducible nuclear pore complex blockade to monitor the kinetics of HIV-1 nuclear import and define the biochemical staging of these steps of infection. Surprisingly, we observe that nuclear import occurs with relatively rapid kinetics (<5 h) and precedes the completion of reverse transcription in target cells, demonstrating that reverse transcription is completed in the nucleus. We also observe that HIV-1 remains susceptible to the capsid-destabilizing compound PF74 following nuclear import, revealing that uncoating is completed in the nucleus. Additionally, we observe that certain capsid mutants are insensitive to a Nup62-mediated nuclear pore complex blockade in cells that potently block infection by wild-type capsid, demonstrating that HIV-1 can use distinct nuclear import pathways during infection. These studies collectively define the spatio-temporal staging of critical steps of HIV-1 infection and provide an experimental system to separate and thereby define the cytoplasmic and nuclear stages of infection by other viruses. HIV-1 reverse transcription is found to be completed in the nucleus of the cell using an HIV-1 nuclear import kinetic assay that takes advantage of a nuclear import blockade method to monitor the kinetics of HIV-1 entry and infection.
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