Identification of an intracellular trafficking and assembly pathway for HIV-1 Gag
Identification of an intracellular trafficking and assembly pathway for HIV-1 Gag
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DOI：
10.1111/j.1398-9219.2006.00428.x
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发表时间：
2006-06-01
期刊：
影响因子：
4.5
通讯作者：
Resh, MD
中科院分区：
文献类型：
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作者：
Perlman, M;Resh, MD
逆转录病毒 Gag 蛋白是膜结合多蛋白，对于病毒样颗粒 (VLP) 的形成是必要且充分的。目前尚不清楚 Gag 如何穿过细胞或如何确定颗粒产生的位置。在这里，我们使用两种技术，双砷/四半胱氨酸 (TC) 标记和放线菌酮块释放，来跟踪新合成的 HIV-1 Gag 的贩运。 Gag 首先出现在细胞质中，在核周簇中积累，短暂地穿过多泡体 (MVB) 样区室，然后移动到质膜 (PM)。活细胞成像证实了 Gag 依次通过这些时间中间体。细胞质钙瞬时升高的诱导增加了 MVB 中 Gag、Gag 组装中间体和 VLP 的量，并导致 VLP 释放急剧增加。这些结果定义了 HIV-1 Gag 的细胞内运输途径，该途径使用核周区室和 MVB 作为运输中间体。我们提出，Gag 与 MVB 样区室关联的调节可调节 HIV-1 出芽和颗粒形成的位点。
Retroviral Gag proteins are membrane-bound polyproteins that are necessary and sufficient for virus-like particle (VLP) formation. It is not known how Gag traffics through the cell or how the site of particle production is determined. Here we use two techniques, biarsenical/tetracysteine (TC) labeling and release from a cycloheximide block, to follow the trafficking of newly synthesized HIV-1 Gag. Gag first appears diffusely distributed in the cytosol, accumulates in perinuclear clusters, passes transiently through a multivesicular body (MVB)-like compartment, and then travels to the plasma membrane (PM). Sequential passage of Gag through these temporal intermediates was confirmed by live cell imaging. Induction of a transient rise in cytoplasmic calcium increased the amounts of Gag, Gag assembly intermediates and VLPs in MVBs, and resulted in a dramatic increase in VLP release. These results define an intracellular trafficking pathway for HIV-1 Gag that uses perinuclear compartments and the MVB as trafficking intermediates. We propose that the regulation of Gag association with MVB-like compartments regulates the site of HIV-1 budding and particle formation.