Aydin Inci, Weber Susanne, Snijder Berend, Samperio Ventayol Pilar, Kühbacher Andreas, Becker Miriam, Day Patricia M, Schiller John T, Kann Michael, Pelkmans Lucas, Helenius Ari, Schelhaas Mario
Emmy-Noether Group: Virus Endocytosis, Institutes of Molecular Virology and Medical Biochemistry, ZMBE, University of Münster, Münster, Germany; Cluster of Excellence EXC1003, Cells in Motion, Münster, Germany.
Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.
PLoS Pathog. 2014 May 29;10(5):e1004162. doi: 10.1371/journal.ppat.1004162. eCollection 2014 May.
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.
采用两步高通量RNA干扰沉默筛选法来鉴定16型人乳头瘤病毒（HPV16）感染过程中所需的宿主细胞因子。对已验证的命中结果进行分析，发现了一组有丝分裂基因，并揭示了一种以前未确定的病毒DNA（vDNA）导入细胞核的机制。在间期细胞中，病毒被内吞，转运至核周区域并脱壳，但vDNA未能导入细胞核。当间期细胞的核膜穿孔时，HPV16感染发生。在有丝分裂期间，vDNA和L2与中期板上的宿主细胞染色质相关联。因此，我们提出HPV16在有丝分裂期间需要核膜破裂，以使vDNA进入核质。这些结果突出了通过RNA干扰筛选发现的基因在病毒感染研究中的价值。此外，HPV16感染过程中所需的细胞功能列表将为未来病毒-宿主细胞相互作用研究提供资源。
