Human papillomavirus (HPV) infection involves complex interactions with the endocytic transport machinery, which ultimately facilitates the entry of the incoming viral genomes into the trans-Golgi network (TGN) and their subsequent nuclear entry during mitosis. The endosomal pathway is a highly dynamic intracellular transport system, which consists of vesicular compartments and tubular extensions, although it is currently unclear whether incoming viruses specifically alter the endocytic machinery. In this study, using MICAL-L1 as a marker for tubulating endosomes, we show that incoming HPV-16 virions induce a profound alteration in global levels of endocytic tubulation. In addition, we also show a critical requirement for the endoplasmic reticulum (ER)-anchored protein VAP in this process. VAP plays an essential role in actin nucleation and endosome-to-Golgi transport. Indeed, the loss of VAP results in a dramatic decrease in the level of endosomal tubulation induced by incoming HPV-16 virions. This is also accompanied by a marked reduction in virus infectivity. In VAP knockdown cells, we see that the defect in virus trafficking occurs after capsid disassembly but prior to localization at the trans-Golgi network, with the incoming virion-transduced DNA accumulating in Vps29/TGN46-positive hybrid vesicles. Taken together, these studies demonstrate that infection with HPV-16 virions induces marked alterations of endocytic transport pathways, some of which are VAP dependent and required for the endosome-to-Golgi transport of the incoming viral L2/DNA complex.IMPORTANCE Human papillomavirus infectious entry involves multiple interactions with the endocytic transport machinery. In this study, we show that incoming HPV-16 virions induce a dramatic increase in endocytic tubulation. This tubulation requires ER-associated VAP, which plays a critical role in ensuring the delivery of cargoes from the endocytic compartments to the trans-Golgi network. Indeed, the loss of VAP blocks HPV infectious entry at a step after capsid uncoating but prior to localization at the trans-Golgi network. These results define a critical role for ER-associated VAP in endocytic tubulation and in HPV-16 infectious entry.

中文翻译：

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人乳头瘤病毒16感染诱导依赖VAP的内胎管。

人乳头瘤病毒（HPV）感染涉及与内吞运输机制的复杂相互作用，这最终促进了传入的病毒基因组进入反高尔基网络（TGN）以及随后在有丝分裂过程中其核进入。内体途径是一种高度动态的细胞内转运系统，由囊泡区室和肾小管延伸组成，尽管目前尚不清楚传入的病毒是否特异性地改变了内吞机制。在这项研究中，使用MICAL-L1作为管形内体的标记物，我们显示传入的HPV-16病毒体可诱导内吞管形体整体水平的深刻改变。此外，在此过程中，我们还显示了对内质网（ER）锚定蛋白VAP的关键要求。VAP在肌动蛋白成核和内体到高尔基体的运输中起着至关重要的作用。确实，VAP的丧失导致由传入的HPV-16病毒体诱导的内体输卵管水平显着降低。这还伴随着病毒感染性的显着降低。在VAP敲低的细胞中，我们看到病毒运输中的缺陷发生在衣壳解体后，但在反转录高尔基体网络定位之前，传入的病毒体转导的DNA积累在Vps29 / TGN46阳性杂交囊泡中。综上所述，这些研究表明，HPV-16病毒体感染可引起内吞转运途径的显着改变，其中某些是VAP依赖性的，是传入病毒L2 / DNA复合体从内体到高尔基体转运的必需条件。重要事项人乳头瘤病毒的感染进入涉及与内吞运输机制的多种相互作用。在这项研究中，我们表明传入的HPV-16病毒体会引起内吞管的急剧增加。这种输卵管需要与ER相关的VAP，这在确保货物从内吞隔室到反高尔基网络的运输中起着至关重要的作用。确实，VAP的丧失会在衣壳解壳后但在反高尔基体网络定位之前的某个步骤中阻止HPV传染性进入。这些结果确定了ER相关的VAP在内吞管和HPV-16感染性进入中的关键作用。在确保将货物从内吞隔室运送到反高尔基网络中起着至关重要的作用。确实，VAP的丧失会在衣壳解壳后但在反穿高尔基体网络定位之前的某个步骤阻止HPV传染性进入。这些结果确定了ER相关的VAP在内吞管和HPV-16感染性进入中的关键作用。在确保将货物从内吞隔室运送到反高尔基网络中起着至关重要的作用。确实，VAP的丧失会在衣壳解壳后但在反高尔基体网络定位之前的某个步骤中阻止HPV传染性进入。这些结果确定了ER相关的VAP在内吞管和HPV-16感染性进入中的关键作用。