HIV-1 cores retain their integrity until minutes before uncoating in the nucleus.
HIV-1 cores retain their integrity until minutes before uncoating in the nucleus.
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DOI：
10.1073/pnas.2019467118
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发表时间：
2021-03-09
影响因子：
11.1
通讯作者：
Pathak VK
中科院分区：
文献类型：
--
作者：
Li C;Burdick RC;Nagashima K;Hu WS;Pathak VK
在这里，我们使用了一种荧光蛋白，该蛋白在溶液中是游离的，并被困在核HIV-1衣壳中，以证明衣壳保持完整性，并防止病毒核心和细胞环境中的大分子混合，直到整合之前。我们还发现，衣壳的完整性保持，直到几分钟前在核中解体，揭示了脱壳后的完整性损失迅速进行。这些有价值的见解到HIV-1复制的早期阶段表明，完整的HIV-1衣壳是通过核孔进口，逆转录主要是完整的衣壳内完成，和整合前复杂的主机相互作用，促进整合和目标位点的选择必须发生在一个很短的时间框架内衣壳解体和整合。我们最近报道，HIV-1核心保留>94%的衣壳（CA）蛋白进入细胞核，并在整合前1.5小时内在整合位点附近解体（未涂层）。然而，核衣壳是否通过破裂或衣壳解体前CA的少量损失而失去其完整性尚不清楚。在这里，我们利用了以前报道的载体，其中绿色荧光蛋白插入HIV-1 Gag（iGFP）;蛋白水解加工有效地释放GFP，其中一些仍然被困在衣壳内，并作为一个流体相内容物标记，当衣壳失去其完整性时释放。我们发现，核衣壳保持其完整性，直到不久前整合，并失去了其GFP含量标志物的mRuby标记的切割和多聚腺苷酸化特异性因子6（mRuby-CPSF 6）的损失前1至3分钟。相比之下，与CA和mRuby-CPSF 6融合的GFP的损失同时发生，表明病毒核心保持其完整性，直到脱壳前几分钟。我们的研究结果表明，HIV-1进化到保持其衣壳的完整性，并保持病毒核心和核环境中的大分子之间的分离，直到脱壳前发生整合。这些观察结果意味着，完整的HIV-1衣壳进口通过核孔;逆转录发生在一个完整的衣壳;和整合前复合物和LEDGF/p75之间的相互作用，可能其他宿主因子，促进整合，必须发生在短时间内损失的衣壳完整性和整合。
Here, we used a fluorescent protein that is free in solution and is trapped in nuclear HIV-1 capsids to demonstrate that the capsids retain integrity and prevent mixing of macromolecules within the viral core and the cellular environment until just before integration. We also found that capsid integrity is maintained until just minutes before disassembly in the nucleus, revealing that uncoating proceeds rapidly after integrity loss. These valuable insights into the early stage of HIV-1 replication indicate that intact HIV-1 capsids are imported through nuclear pores, that reverse transcription is mostly completed within intact capsids, and that preintegration complex-host interactions facilitating integration and target site selection must occur within a short time frame between capsid disassembly and integration. We recently reported that HIV-1 cores that retained >94% of their capsid (CA) protein entered the nucleus and disassembled (uncoated) near their integration site <1.5 h before integration. However, whether the nuclear capsids lost their integrity by rupturing or a small loss of CA before capsid disassembly was unclear. Here, we utilized a previously reported vector in which green fluorescent protein is inserted in HIV-1 Gag (iGFP); proteolytic processing efficiently releases GFP, some of which remains trapped inside capsids and serves as a fluid phase content marker that is released when the capsids lose their integrity. We found that nuclear capsids retained their integrity until shortly before integration and lost their GFP content marker ∼1 to 3 min before loss of capsid-associated mRuby-tagged cleavage and polyadenylation specificity factor 6 (mRuby-CPSF6). In contrast, loss of GFP fused to CA and mRuby-CPSF6 occurred simultaneously, indicating that viral cores retain their integrity until just minutes before uncoating. Our results indicate that HIV-1 evolved to retain its capsid integrity and maintain a separation between macromolecules in the viral core and the nuclear environment until uncoating occurs just before integration. These observations imply that intact HIV-1 capsids are imported through nuclear pores; that reverse transcription occurs in an intact capsid; and that interactions between the preintegration complex and LEDGF/p75, and possibly other host factors that facilitate integration, must occur during the short time period between loss of capsid integrity and integration.
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DOI：
10.1038/nrmicro3503
发表时间：
2015-08
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
Campbell EM;Hope TJ
通讯作者：
Hope TJ
影响因子：
4.8
作者：
Cherepanov, P;Maertens, G;Debyser, Z
通讯作者：
Debyser, Z
影响因子：
5.4
作者：
Balasubramaniam, Muthukumar;Zhou, Jing;Dash, Chandravanu
通讯作者：
Dash, Chandravanu
影响因子：
7.7
作者：
Bejarano, David Alejandro;Peng, Ke;Kraeusslich, Hans-Georg
通讯作者：
Kraeusslich, Hans-Georg
DOI：
10.1073/pnas.1419945112
发表时间：
2014-12-30
影响因子：
11.1
作者：
Bhattacharya, Akash;Alam, Steven L.;Yeager, Mark
通讯作者：
Yeager, Mark