The Role of Protein Kinase A Regulation of the E6 PDZ-Binding Domain during the Differentiation-Dependent Life Cycle of Human Papillomavirus Type 18
Open Access
- 1 September 2013
- journal article
- research article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 87 (17), 9463-9472
- https://doi.org/10.1128/jvi.01234-13
Abstract
Human papillomavirus (HPV) E6 proteins of high-risk alpha types target a select group of PSD95/DLG1/ZO1 (PDZ) domain-containing proteins by using a C-terminal PDZ-binding motif (PBM), an interaction that can be negatively regulated by phosphorylation of the E6 PBM by protein kinase A (PKA). Here, we have mutated the canonical PKA recognition motif that partially overlaps with the E6 PBM in the HPV18 genome (E6153PKA) and compared the effect of this mutation on the HPVl8 life cycle in primary keratinocytes with the wild-type genome and with a second mutant genome that lacks the E6 PBM (E6ΔPDZ). Loss of PKA recognition of E6 was associated with increased growth of the genome-containing cells relative to cells carrying the wild-type genome, and upon stratification, a more hyperplastic phenotype, with an increase in the number of S-phase competent cells in the upper suprabasal layers, while the opposite was seen with the E6ΔPDZ genome. Moreover, the growth of wild-type genome-containing cells was sensitive to changes in PKA activity, and these changes were associated with increased phosphorylation of the E6 PBM. In marked contrast to E6ΔPDZ genomes, the E6153PKA mutation exhibited no deleterious effects on viral genome amplification or expression of late proteins. Our data suggest that the E6 PBM function is differentially regulated by phosphorylation in the HPV18 life cycle. We speculate that perturbation of protein kinase signaling pathways could lead to changes in E6 PBM function, which in turn could have a bearing on tumor promotion and progression.Keywords
This publication has 42 references indexed in Scilit:
- Human papillomavirus genotype attribution in invasive cervical cancer: a retrospective cross-sectional worldwide studyThe Lancet Oncology, 2010
- The Human Papillomavirus (HPV) E6* Proteins from High-Risk, Mucosal HPVs Can Direct Degradation of Cellular Proteins in the Absence of Full-Length E6 ProteinJournal of Virology, 2009
- Disruption of the E2 Gene Is a Common and Early Event in the Natural History of Cervical Human Papillomavirus Infection: A Longitudinal Cohort StudyCancer Research, 2009
- Robust production and passaging of infectious HPV in squamous epithelium of primary human keratinocytesGenes & Development, 2009
- The PDZ Binding Motif of Human Papillomavirus Type 16 E6 Induces PTPN13 Loss, Which Allows Anchorage-Independent Growth and Synergizes with Ras for Invasive GrowthJournal of Virology, 2008
- Human Papillomavirus Types in Head and Neck Squamous Cell Carcinomas Worldwide: A Systematic ReviewCancer Epidemiology, Biomarkers & Prevention, 2005
- Requirement of PDZ-Containing Proteins for Cell Cycle Regulation and Differentiation in the Mouse Lens EpitheliumMolecular and Cellular Biology, 2003
- The PDZ Ligand Domain of the Human Papillomavirus Type 16 E6 Protein Is Required for E6's Induction of Epithelial Hyperplasia In VivoJournal of Virology, 2003
- Oncogenic human papillomavirus E6 proteins target the MAGI-2 and MAGI-3 proteins for degradationOncogene, 2002
- Human papillomavirus type 31 oncoproteins E6 and E7 are required for the maintenance of episomes during the viral life cycle in normal human keratinocytesProceedings of the National Academy of Sciences, 1999