HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines and in Primary Macrophages.
HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines and in Primary Macrophages.
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DOI：
10.3390/v12111234
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发表时间：
2020-10-30
期刊：
影响因子：
--
通讯作者：
Melikyan GB
中科院分区：
文献类型：
--
作者：
Francis AC;Marin M;Prellberg MJ;Palermino-Rowland K;Melikyan GB
衣壳蛋白（CA）的组装形成包围HIV-1核糖核蛋白复合物的成熟病毒核心。关于感染细胞中的细胞位点和核心解体（未包被）的程度，已报告了不一致的发现。在这里，我们结合单病毒成像和药物添加时间测定来阐明细胞系和单核细胞衍生的巨噬细胞（MDM）中HIV-1复合物的脱壳，逆转录和核输入之间的动力学关系。通过使用亲环素A-DsRed（CDR）作为CA的标记物，我们表明，与TZM-bl细胞相反，MDM中早期细胞质未包被（CDR的损失）是有限的，并且与逆转录的效率相关。然而，我们发现逆转录是HIV-1核输入，这是通过在核孔的脱壳步骤进展。核输入的动力学和病毒从针对感染的不同步骤的抑制剂逃逸的比较，以及病毒DNA合成的直接定量，揭示了HIV-1复合物的核输入后完成逆转录。总的来说，这些结果表明，逆转录是在核孔的脱壳步骤，并在不相关的靶细胞的细胞核中完成vDNA合成。
An assembly of capsid proteins (CA) form the mature viral core enclosing the HIV-1 ribonucleoprotein complex. Discrepant findings have been reported regarding the cellular sites and the extent of core disassembly (uncoating) in infected cells. Here, we combined single-virus imaging and time-of-drug-addition assays to elucidate the kinetic relationship between uncoating, reverse transcription, and nuclear import of HIV-1 complexes in cell lines and monocyte-derived macrophages (MDMs). By using cyclophilin A-DsRed (CDR) as a marker for CA, we show that, in contrast to TZM-bl cells, early cytoplasmic uncoating (loss of CDR) is limited in MDMs and is correlated with the efficiency of reverse transcription. However, we find that reverse transcription is dispensable for HIV-1 nuclear import, which progressed through an uncoating step at the nuclear pore. Comparison of the kinetics of nuclear import and the virus escape from inhibitors targeting distinct steps of infection, as well as direct quantification of viral DNA synthesis, revealed that reverse transcription is completed after nuclear import of HIV-1 complexes. Collectively, these results suggest that reverse transcription is dispensable for the uncoating step at the nuclear pore and that vDNA synthesis is completed in the nucleus of unrelated target cells.
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DOI：
10.1038/nrmicro3503
发表时间：
2015-08
期刊：
Nature reviews. Microbiology
影响因子：
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作者：
Campbell EM;Hope TJ
通讯作者：
Hope TJ
影响因子：
5.4
作者：
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影响因子：
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通讯作者：
Kraeusslich, Hans-Georg
DOI：
10.1073/pnas.1700247114
发表时间：
2017-08-15
影响因子：
11.1
作者：
Delaney, Michael Keegan;Malikov, Viacheslav;Naghavi, Mojgan H.
通讯作者：
Naghavi, Mojgan H.
DOI：
10.1073/pnas.1419945112
发表时间：
2014-12-30
影响因子：
11.1
作者：
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通讯作者：
Yeager, Mark