Large scale RNAi reveals the requirement of nuclear envelope breakdown for nuclear import of human papillomaviruses

PLoS Pathog. 2014 May 29;10(5):e1004162. doi: 10.1371/journal.ppat.1004162. eCollection 2014 May.

Abstract

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.

MeSH terms

  • Active Transport, Cell Nucleus
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • DNA, Viral / genetics
  • Human papillomavirus 16* / genetics
  • Humans
  • Mitosis / physiology*
  • Nuclear Envelope / metabolism*
  • Oncogene Proteins, Viral / genetics*
  • RNA Interference*

Substances

  • DNA, Viral
  • Oncogene Proteins, Viral

Grants and funding

MS was supported by the German Research Foundation (DFG, grants SCHE 1552/2-1, GRK 1409/C2, EXC 1003 (partly)). AH was supported by the ETH Zurich, and by grants from the Swiss National Science Foundation (SNF) and the European Research Council (ERC). LP was supported by the ETH Zurich, the University of Zurich, the European Union, and by SystemsX.ch, the Swiss Initiative in Systems Biology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.