Vesicular trafficking of incoming human papillomavirus 16 to the Golgi apparatus and endoplasmic reticulum requires γ-secretase activity.
Vesicular trafficking of incoming human papillomavirus 16 to the Golgi apparatus and endoplasmic reticulum requires γ-secretase activity.
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DOI：
10.1128/mbio.01777-14
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发表时间：
2014-09-16
期刊：
影响因子：
6.4
通讯作者：
DiMaio D
中科院分区：
文献类型：
--
作者：
Zhang W;Kazakov T;Popa A;DiMaio D
乳头瘤病毒在感染过程中从细胞表面到细胞核的途径还不完全清楚。在这里，我们开发了一种新的人乳头瘤病毒16（HPV 16）假病毒，其中的次要衣壳蛋白L2的羧基末端暴露在细胞结合之前的完整衣壳的外部。对于这种假病毒，我们使用邻近连接试验免疫检测技术来证明，在进入期间，HPV 16 L2在高尔基体定位之前进出早期内体，并且我们证明了L2在进入期间进入内质网。细胞膜相关蛋白酶γ-分泌酶是HPV 16假病毒和真HPV 16感染所必需的。我们还发现，γ-分泌酶的抑制不会实质性地干扰病毒内化、衣壳解体的启动、进入早期核内体或从该区室退出，但γ-分泌酶是L2和病毒DNA定位于高尔基体和内质网所必需的。这些结果表明，传入的HPV 16运输顺序从细胞表面到内体，然后到高尔基体和内质网，然后进入核。人乳头瘤病毒是小型无包膜DNA病毒，约占所有人类癌症死亡的5%，但对这些病毒从细胞表面转移到细胞核的过程知之甚少。在这里，我们表明，传入的HPV 16，最常见的高危型HPV，交通通过一系列的囊泡隔间在感染性进入，包括核内体，高尔基体，和内质网。此外，我们表明，γ-分泌酶，细胞膜相关的蛋白酶，需要的L2小衣壳蛋白和病毒DNA进入高尔基体和内质网。这些研究揭示了DNA病毒进入细胞的新途径，并表明该途径的组分是候选的抗病毒靶点。
The route taken by papillomaviruses from the cell surface to the nucleus during infection is incompletely understood. Here, we developed a novel human papillomavirus 16 (HPV16) pseudovirus in which the carboxy terminus of the minor capsid protein L2 is exposed on the exterior of the intact capsid prior to cell binding. With this pseudovirus, we used the proximity ligation assay immune detection technique to demonstrate that during entry HPV16 L2 traffics into and out of the early endosome prior to Golgi localization, and we demonstrated that L2 enters the endoplasmic reticulum during entry. The cellular membrane-associated protease, γ-secretase, is required for infection by HPV16 pseudovirus and authentic HPV16. We also showed that inhibition of γ-secretase does not interfere substantively with virus internalization, initiation of capsid disassembly, entry into the early endosome, or exit from this compartment, but γ-secretase is required for localization of L2 and viral DNA to the Golgi apparatus and the endoplasmic reticulum. These results show that incoming HPV16 traffics sequentially from the cell surface to the endosome and then to the Golgi apparatus and the endoplasmic reticulum prior to nuclear entry. The human papillomaviruses are small nonenveloped DNA viruses responsible for approximately 5% of all human cancer deaths, but little is known about the process by which these viruses transit from the cell surface to the nucleus. Here we show that incoming HPV16, the most common high-risk HPV, traffics though a series of vesicular compartments during infectious entry, including the endosome, Golgi apparatus, and endoplasmic reticulum. Furthermore, we show that γ-secretase, a cellular membrane-associated protease, is required for entry of the L2 minor capsid protein and viral DNA into the Golgi apparatus and endoplasmic reticulum. These studies reveal a new pathway of cell entry by DNA viruses and suggest that components of this pathway are candidate antiviral targets.