Compromised spindle assembly checkpoint due to altered expression of Ubch10 and Cdc20 in human papillomavirus type 16 E6- and E7-expressing keratinocytes

J Virol. 2010 Nov;84(21):10956-64. doi: 10.1128/JVI.00259-10. Epub 2010 Aug 25.

Abstract

Cells expressing human papillomavirus type 16 (HPV-16) E6 and E7 proteins exhibit deregulation of G2/M genes, allowing bypass of DNA damage arrest signals. Normally, cells with DNA damage that override the G2 damage checkpoint would precociously enter mitosis and ultimately face mitotic catastrophe and apoptotic cell death. However, E6/E7-expressing cells (E6/E7 cells) have the ability to enter and exit mitosis in the presence of DNA damage and continue with the next round of the cell cycle. Little is known about the mechanism that allows these cells to gain entry into and exit from mitosis. Here, we show that in the presence of DNA damage, E6/E7 cells have elevated levels of cyclin B, which would allow entry into mitosis. Also, as required for exit from mitosis, cyclin B is degraded in these cells, permitting initiation of the next round of DNA synthesis and cell cycle progression. Proteasomal degradation of cyclin B by anaphase-promoting complex/cyclosome (APC/C) is, in part, due to elevated levels of the E2-conjugating enzyme, Ubch10, and the substrate recognition protein, Cdc20, of APC/C. Also, in E6/E7 cells with DNA damage, while Cdc20 is complexed with BubR1, indicating an active checkpoint, it is also present in complexes free of BubR1, presumably allowing APC/C activity and slippage through the checkpoint.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cdc20 Proteins
  • Cell Cycle
  • Cell Cycle Proteins / genetics*
  • Cyclin B / metabolism
  • DNA Damage
  • Gene Expression Regulation*
  • Human papillomavirus 16 / physiology*
  • Humans
  • Keratinocytes / metabolism
  • Keratinocytes / virology*
  • Mitosis
  • Oncogene Proteins, Viral / genetics*
  • Papillomavirus E7 Proteins / genetics*
  • Repressor Proteins / genetics*
  • Spindle Apparatus / pathology
  • Spindle Apparatus / virology*
  • Ubiquitin-Conjugating Enzymes / genetics*

Substances

  • Cdc20 Proteins
  • Cell Cycle Proteins
  • Cyclin B
  • E6 protein, Human papillomavirus type 16
  • Oncogene Proteins, Viral
  • Papillomavirus E7 Proteins
  • Repressor Proteins
  • oncogene protein E7, Human papillomavirus type 16
  • CDC20 protein, human
  • UBE2C protein, human
  • Ubiquitin-Conjugating Enzymes