HIV-1 Capsid Uncoating Is a Multistep Process That Proceeds through Defect Formation Followed by Disassembly of the Capsid Lattice.
HIV-1 Capsid Uncoating Is a Multistep Process That Proceeds through Defect Formation Followed by Disassembly of the Capsid Lattice.
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HIV-1衣壳脱壳是一个多步骤的过程，通过缺陷的形成，然后衣壳晶格的破坏。
DOI：
10.1021/acsnano.3c07678
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发表时间：
2024-01-30
期刊：
影响因子：
17.1
通讯作者：
Melikyan, Gregory B.
中科院分区：
文献类型：
--
作者：
Gifford, Levi B.;Melikyan, Gregory B.
关键词：
HIV-1 核心由锥形衣壳组成，该衣壳由衣壳蛋白 (CA) 六聚体和五聚体组成，封装病毒基因组。 HIV-1 衣壳解体（称为脱衣）对于生产性感染非常重要；然而，脱涂层的位置、时间和规定仍然存在争议。在这里，我们采用琥珀密码子抑制来直接标记 CA。此外，将液相荧光探针整合到病毒核心中以检测衣壳晶格中的小缺陷。这种双标记策略能够在体外和活细胞中实现单核脱壳的可视化，我们发现这总是通过至少两个不同的步骤进行——在衣壳晶格中形成缺陷，从而导致 CA 逐渐损失到可检测水平以下。重要的是，在建立有效感染之前，含有液相和 CA 荧光标记的完整核心进入细胞核并在细胞核中脱壳，这可以通过两种标记的连续损失来证明。在不同的细胞（包括巨噬细胞系）中观察到这种两步脱壳过程。值得注意的是，液相标记物的释放和 CA 的最终丢失之间的滞后似乎与细胞类型或逆转录无关，并且与无细胞核心或细胞质中的核心相比，核衣壳的滞后时间要长得多（> 5 倍），这表明衣壳晶格是通过衣壳结合核因子稳定的。我们的结果表明，完整的 HIV-1 核心进入细胞核，并且在 CA 整体丧失之前，通过衣壳晶格中的局部缺陷启动脱壳。
The HIV-1 core consists of a cone-shaped capsid shell made of capsid protein (CA) hexamers and pentamers encapsulating the viral genome. HIV-1 capsid disassembly, referred to as uncoating, is important for productive infection; however, the location, timing, and regulation of uncoating remain controversial. Here, we employ amber codon suppression to directly label CA. In addition, a fluid phase fluorescent probe is incorporated into the viral core to detect small defects in the capsid lattice. This double-labeling strategy enables the visualization of uncoating of single cores in vitro and in living cells, which we found to always proceed through at least two distinct steps—the formation of a defect in the capsid lattice that initiates gradual loss of CA below a detectable level. Importantly, intact cores containing the fluid phase and CA fluorescent markers enter and uncoat in the nucleus, as evidenced by a sequential loss of both markers, prior to establishing productive infection. This two-step uncoating process is observed in different cells, including a macrophage line. Notably, the lag between the release of fluid phase marker and terminal loss of CA appears to be independent of the cell type or reverse transcription and is much longer (>5-fold) for nuclear capsids compared to cell-free cores or cores in the cytosol, suggesting that the capsid lattice is stabilized by capsid-binding nuclear factors. Our results imply that intact HIV-1 cores enter the cell nucleus and that uncoating is initiated through a localized defect in the capsid lattice prior to a global loss of CA.
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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：
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通讯作者：
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影响因子：
3.7
作者：
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影响因子：
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作者：
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Dash, Chandravanu
影响因子：
3.7
作者：
Bonifati S;Daly MB;St Gelais C;Kim SH;Hollenbaugh JA;Shepard C;Kennedy EM;Kim DH;Schinazi RF;Kim B;Wu L
通讯作者：
Wu L
DOI：
10.3390/v12111234
发表时间：
2020-10-30
期刊：
Viruses
影响因子：
--
作者：
Francis AC;Marin M;Prellberg MJ;Palermino-Rowland K;Melikyan GB
通讯作者：
Melikyan GB