Large scale RNAi reveals the requirement of nuclear envelope breakdown for nuclear import of human papillomaviruses.
Large scale RNAi reveals the requirement of nuclear envelope breakdown for nuclear import of human papillomaviruses.
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DOI：
10.1371/journal.ppat.1004162
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发表时间：
2014-05
期刊：
影响因子：
6.7
通讯作者：
Schelhaas M
中科院分区：
文献类型：
--
作者：
Aydin I;Weber S;Snijder B;Samperio Ventayol P;Kühbacher A;Becker M;Day PM;Schiller JT;Kann M;Pelkmans L;Helenius A;Schelhaas M
采用两步高通量RNAi沉默筛选方法鉴定人乳头瘤病毒16型（HPV16）感染过程中所需的宿主细胞因子。对验证命中的分析涉及一组有丝分裂基因，并揭示了一种以前未确定的病毒DNA （vDNA）进入细胞核的机制。在间期细胞中，病毒被内吞，被输送到核周区域，并被剥去包被，但vDNA不能被输入细胞核。间期细胞核膜穿孔后发生HPV16感染。在有丝分裂过程中，vDNA和L2在中期板上与宿主细胞染色质结合。因此，我们提出HPV16需要在有丝分裂过程中破坏核膜以使vDNA进入核质。结果强调了通过RNAi筛选发现的基因在病毒感染调查中的价值。此外，HPV16感染过程中所需的细胞功能列表将为未来病毒-宿主细胞相互作用的研究提供资源。某些人乳头瘤病毒（HPV）是宫颈癌和其他上皮性肿瘤的病因。抗人乳头瘤病毒疫苗的最新进展及其越来越多的应用为未来这些癌症的显著减少提供了希望。然而，HPV在感染者和未感染者之间传播的许多细节仍然没有得到很好的了解。特别是，进入宿主细胞的多步骤过程仅部分表征。在这里，我们使用了一种系统的RNAi沉默方法来鉴定HPV16感染期间所需的宿主细胞蛋白，HPV16是最常见的致癌HPV。我们的结果表明，HPV的进入需要细胞分裂。细胞分裂很重要，因为它消除了核质和细胞质之间的屏障，这样病毒就可以进入细胞核进行转录和复制。我们的研究为一种重要病原体的进入过程提供了新的机制见解，并解释了为什么它只能感染人类皮肤或粘膜的干细胞或短暂扩增细胞。这项工作还强调了DNA病毒的一种新的核进口策略，它通常使用核进口机制进入核质进行感染，因此，HPV的进入显示了大多数其他病毒不常见的另一个独特方面。
A two-step, high-throughput RNAi silencing screen was used to identify host cell factors required during human papillomavirus type 16 (HPV16) infection. Analysis of validated hits implicated a cluster of mitotic genes and revealed a previously undetermined mechanism for import of the viral DNA (vDNA) into the nucleus. In interphase cells, viruses were endocytosed, routed to the perinuclear area, and uncoated, but the vDNA failed to be imported into the nucleus. Upon nuclear envelope perforation in interphase cells HPV16 infection occured. During mitosis, the vDNA and L2 associated with host cell chromatin on the metaphase plate. Hence, we propose that HPV16 requires nuclear envelope breakdown during mitosis for access of the vDNA to the nucleoplasm. The results accentuate the value of genes found by RNAi screens for investigation of viral infections. The list of cell functions required during HPV16 infection will, moreover, provide a resource for future virus-host cell interaction studies. Certain human papillomaviruses (HPV) are the etiological cause of cervical cancers and other epithelial tumors. Recent advances in the development of anti-HPV vaccines and their increasing deployment provide hope for a significant decrease of these cancers in the future. However, many details of the transmission of HPV between infected and uninfected individuals are still not well understood. In particular, the multistep process of entry into host cells is only partially characterized. Here, we used a systematic RNAi silencing approach to identify host cell proteins required during HPV16 infection, the most prevalent cancer-causing HPV. Our results indicated that HPV entry requires cells to divide. Cell division is important, since it removes the barrier between the nucleoplasm and the cytosol, so that the virus can access the nucleus for transcription and replication. Our study provides new mechanistic insights into the entry process of an important pathogen, and explains why it can only infect the stem cells or transiently amplifying cells of human skin or mucosa. This work also highlights a novel nuclear import strategy for DNA viruses, which typically use the nuclear import machinery to access the nucleoplasm for infection, Thus, HPV entry displays another unique aspect uncommon to most other viruses.
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肝素增加了独立于细胞表面蛋白聚糖的16型人乳头瘤病毒的感染性，并诱导L1表位暴露。
影响因子：
3.4
作者：
Cerqueira C;Liu Y;Kühling L;Chai W;Hafezi W;van Kuppevelt TH;Kühn JE;Feizi T;Schelhaas M
通讯作者：
Schelhaas M
影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
5.4
作者：
Florin, Luise;Becker, Katrin A.;Sapp, Martin
通讯作者：
Sapp, Martin
影响因子：
3.7
作者：
Day, PM;Lowy, DR;Schiller, JT
通讯作者：
Schiller, JT
影响因子：
3.7
作者：
Drobni, P;Mistry, N;Evander, M
通讯作者：
Evander, M