Clathrin facilitates the morphogenesis of retrovirus particles.
Clathrin facilitates the morphogenesis of retrovirus particles.
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DOI：
10.1371/journal.ppat.1002119
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发表时间：
2011-06
期刊：
影响因子：
6.7
通讯作者：
Bieniasz PD
中科院分区：
文献类型：
--
作者：
Zhang F;Zang T;Wilson SJ;Johnson MC;Bieniasz PD
逆转录病毒颗粒的形态发生由 Gag 和 GagPol 蛋白驱动，它们提供颗粒组装和成熟所需的主要结构成分和酶活性。此外，在逆转录病毒颗粒中还发现了许多细胞蛋白；其中一些对于病毒复制很重要，但许多缺乏已知的功能作用。其中一种蛋白质是网格蛋白，由于其在质膜上的丰富性，人们认为它会被动地掺入病毒粒子中。我们发现，网格蛋白不仅在高度纯化的 HIV-1 颗粒中异常丰富，而且以高特异性招募。特别是，HIV-1 Pol 蛋白是网格蛋白掺入所必需的，并且 Pol 的逆转录酶或整合酶结构域中的点突变可能会消除掺入。网格蛋白还被特异性地掺入其他逆转录病毒颗粒中，包括慢病毒（猿猴免疫缺陷病毒，SIVmac）、γ逆转录病毒（鼠白血病病毒，MLV）和β逆转录病毒（梅森-辉瑞猴病毒，M-PMV）属的成员。然而，与 HIV-1 不同的是，这些其他逆转录病毒主要利用各自 Gag 蛋白中的肽基序来招募网格蛋白，这些肽基序模仿了细胞网格蛋白接头中发现的基序。通过病毒蛋白诱变、基于 siRNA 的网格蛋白耗竭或接头蛋白 (AP180) 诱导的网格蛋白隔离，干扰网格蛋白掺入这些逆转录病毒，对颗粒形态发生的准确性产生一系列影响。这些影响根据所检测的逆转录病毒而异，包括 Gag 和/或 Pol 蛋白不稳定、颗粒组装抑制和病毒粒子感染性降低。对于所检查的每种逆转录病毒，网格蛋白的掺入似乎对于最佳复制很重要。这些数据表明许多逆转录病毒利用网格蛋白来促进感染性颗粒的准确形态发生。我们提出了一个模型，其中网格蛋白有助于 Gag 和 Pol 蛋白的空间组织，从而调节颗粒组装过程中病毒体成分的蛋白水解加工。传染性逆转录病毒的组装和成熟是由两种病毒蛋白 Gag 和 Pol 驱动的。此外，在逆转录病毒颗粒中发现了许多细胞蛋白，其中许多缺乏已知的功能作用。其中一种蛋白质是网格蛋白，它通常介导细胞中的多种生理过程，以前被认为只是被动地掺入病毒粒子中。在这项研究中，我们表明网格蛋白主动、特异且大量地融入逆转录病毒颗粒中。在一些情况下，逆转录病毒蛋白编码的肽基序模仿了负责网格蛋白募集的细胞衔接蛋白中发现的肽基序。包装网格蛋白的逆转录病毒范围包括人类和猿类免疫缺陷病毒以及其他鼠类和猿类逆转录病毒。阻止网格蛋白掺入病毒颗粒的操作也会导致感染性逆转录病毒的发生过程中出现各种缺陷，包括病毒蛋白不稳定、颗粒组装和释放的抑制以及病毒颗粒传染性的降低。缺陷的确切性质根据所检查的特定逆转录病毒而有所不同。总的来说，这些研究表明网格蛋白经常被逆转录病毒用来促进感染性病毒颗粒的准确组装。
The morphogenesis of retroviral particles is driven by Gag and GagPol proteins that provide the major structural component and enzymatic activities required for particle assembly and maturation. In addition, a number of cellular proteins are found in retrovirus particles; some of these are important for viral replication, but many lack a known functional role. One such protein is clathrin, which is assumed to be passively incorporated into virions due to its abundance at the plasma membrane. We found that clathrin is not only exceptionally abundant in highly purified HIV-1 particles but is recruited with high specificity. In particular, the HIV-1 Pol protein was absolutely required for clathrin incorporation and point mutations in reverse transcriptase or integrase domains of Pol could abolish incorporation. Clathrin was also specifically incorporated into other retrovirus particles, including members of the lentivirus (simian immunodeficiency virus, SIVmac), gammaretrovirus (murine leukemia virus, MLV) and betaretrovirus (Mason-Pfizer monkey virus, M-PMV) genera. However, unlike HIV-1, these other retroviruses recruited clathrin primarily using peptide motifs in their respective Gag proteins that mimicked motifs found in cellular clathrin adaptors. Perturbation of clathrin incorporation into these retroviruses, via mutagenesis of viral proteins, siRNA based clathrin depletion or adaptor protein (AP180) induced clathrin sequestration, had a range of effects on the accuracy of particle morphogenesis. These effects varied according to which retrovirus was examined, and included Gag and/or Pol protein destabilization, inhibition of particle assembly and reduction in virion infectivity. For each retrovirus examined, clathrin incorporation appeared to be important for optimal replication. These data indicate that a number of retroviruses employ clathrin to facilitate the accurate morphogenesis of infectious particles. We propose a model in which clathrin contributes to the spatial organization of Gag and Pol proteins, and thereby regulates proteolytic processing of virion components during particle assembly. The assembly and maturation of infectious retroviruses is driven by two viral proteins, Gag and Pol. Additionally, a number of cellular proteins are found in retrovirus particles, many of which lack a known functional role. One such protein is clathrin, which normally mediates several physiological processes in cells and was previously thought to be only passively incorporated into virions. In this study we show that clathrin is actively, specifically and abundantly incorporated into retrovirus particles. In several cases, retroviral proteins encode peptide motifs that mimic those found in cellular adaptor proteins that are responsible for clathrin recruitment. The range of retroviruses into which clathrin is packaged includes human and simian immunodeficiency viruses as well as other murine and simian retroviruses. Manipulations that prevented clathrin incorporation into virions also caused a variety of defects in the genesis of infectious retroviruses, including viral protein destabilization, inhibition of particle assembly and release, and reduction in virion infectiousness. The precise nature of the defect varied according to which particular retrovirus was examined. Overall these studies suggest that clathrin is frequently employed by retroviruses to facilitate the accurate assembly of infectious virions.
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影响因子：
4.8
作者：
Jaeger, Stefanie;Gulbahce, Natali;Krogan, Nevan J.
通讯作者：
Krogan, Nevan J.
DOI：
10.1083/jcb.200408155
发表时间：
2005-01-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Martin-Serrano J;Eastman SW;Chung W;Bieniasz PD
通讯作者：
Bieniasz PD
影响因子：
3.7
作者：
Chiang, Chien-Cheng;Wang, Shiu-Mei;Wang, Chin-Tien
通讯作者：
Wang, Chin-Tien
影响因子：
5.4
作者：
Müller, B;Daecke, J;Kräusslich, HG
通讯作者：
Kräusslich, HG
影响因子：
5.4
作者：
Lu, R;Limón, A;Engelman, A
通讯作者：
Engelman, A