Anna Severino and Claudia Abbruzzese contributed equally to this work.
Human papillomavirus-16 E7 interacts with siva-1 and modulates apoptosis in HaCaT human immortalized keratinocytes†
Article first published online: 8 MAR 2007
Copyright © 2007 Wiley-Liss, Inc.
Journal of Cellular Physiology
Volume 212, Issue 1, pages 118–125, July 2007
How to Cite
Severino, A., Abbruzzese, C., Manente, L., Valderas, Á. A., Mattarocci, S., Federico, A., Starace, G., Chersi, A., Mileo, A. M. and Paggi, M. G. (2007), Human papillomavirus-16 E7 interacts with siva-1 and modulates apoptosis in HaCaT human immortalized keratinocytes. J. Cell. Physiol., 212: 118–125. doi: 10.1002/jcp.21011
- Issue published online: 26 APR 2007
- Article first published online: 8 MAR 2007
- Manuscript Accepted: 5 DEC 2006
- Manuscript Received: 31 OCT 2006
- Associazione Italiana Ricerca sul Cancro
- Ministero della Salute
- Marie Curie Training Site. Grant Number: HPMT-CT-2000-00210
The viral factor E7 plays a key role in the well-established association between “high-risk” Human Papillomavirus (HPV) infection and the development of epithelial malignant tumors, as uterine cervix and ano-genital cancer. To delve into the molecular mechanisms of HPV-mediated cell transformation, we searched for novel potential cellular targets of the HPV-16 E7 oncoprotein, by means of the yeast two-hybrid technique, identifying a protein–protein interaction between HPV-16 E7 and the pro-apoptotic cellular factor Siva-1. Using co-precipitation assays and the “PepSets” technique, we confirmed this physical interaction and mapped accurately, for both proteins, the amino acid residues involved. Additionally, we found that HPV-16 E7 competed in vitro with the binding of the Bcl-XL anti-apoptotic factor to Siva-1, an interaction that has a major inference in UV radiation-induced apoptosis. In HaCaT immortalized human keratinocytes, forced HPV-16 E7 expression by retroviral infection caused Siva-1 transcript up-regulation, detected by cDNA macroarray hybridization and real-time quantitative PCR, paralleled by an increased amount of protein. Confirming the anti-apoptotic role of HPV-16 E7 in the HaCaT cellular model, evaluated by nuclear morphology, we also found that Siva-1 expression produced a significant increase of the apoptotic rate in UV radiation-exposed HaCaT cells, and that this effect resulted explicitly counteracted by HPV-16 E7. Being apoptosis a key physiological process for the elimination of irreversibly injured cells, the anti-apoptotic role of HPV-16 E7, performed at least by its interference with Siva-1, can be considered an additional mechanism for the survival of damaged, potentially transforming, cell clones. J. Cell. Physiol. 212: 118–125, 2007. © 2007 Wiley-Liss, Inc.