Each papillomavirus is species specific and replicates persistently in a specific type of cutaneous or mucosal epithelium. The keratinocytes of the basal layer of the epithelium harbor a reservoir of replicating viral genomes and therefore are the ideal cells in which to study the mechanisms by which HPV establishes persistent infection. Papillomaviruses will only undergo their complete life cycle and generate progeny virus in the tissue culture equivalent of a stratified epithelium. This process requires establishing stably replicating viral DNA genomes in primary human keratinocytes and culturing them in three-dimensional skin equivalents. For our studies on HPV replication we wanted to develop an optimal replication system in keratinocytes so that we could study the requirements in the natural host. Alpha Papillomaviruses induce persistent epithelial lesions and are associated with the development of cervical cancer.
The aims of this project are to determine the mechanism of extrachromosomal replication and partitioning of the viral genome. -We have established a treatment that allows us to efficiently immortalize primary human keratinocytes. -The immortalized keratinocytes are invaluable for studies on the long-term persistent viral DNA replication

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2010
Total Cost
$177,503
Indirect Cost
City
State
Country
Zip Code
Porter, Samuel S; Stepp, Wesley H; Stamos, James D et al. (2017) Host cell restriction factors that limit transcription and replication of human papillomavirus. Virus Res 231:10-20
McBride, Alison A (2017) Mechanisms and strategies of papillomavirus replication. Biol Chem 398:919-927
Van Doorslaer, Koenraad; Chen, Dan; Chapman, Sandra et al. (2017) Persistence of an Oncogenic Papillomavirus Genome Requires cis Elements from the Viral Transcriptional Enhancer. MBio 8:
McBride, Alison A (2017) Playing with fire: consequences of human papillomavirus DNA replication adjacent to genetically unstable regions of host chromatin. Curr Opin Virol 26:63-68
Dooley, Katharine E; Warburton, Alix; McBride, Alison A (2016) Tandemly Integrated HPV16 Can Form a Brd4-Dependent Super-Enhancer-Like Element That Drives Transcription of Viral Oncogenes. MBio 7:
Van Doorslaer, Koenraad; Porter, Samuel; McKinney, Caleb et al. (2016) Novel recombinant papillomavirus genomes expressing selectable genes. Sci Rep 6:37782
McKinney, Caleb C; Kim, Min Jung; Chen, Dan et al. (2016) Brd4 Activates Early Viral Transcription upon Human Papillomavirus 18 Infection of Primary Keratinocytes. MBio 7:
Sakakibara, Nozomi; Chen, Dan; McBride, Alison A (2013) Papillomaviruses use recombination-dependent replication to vegetatively amplify their genomes in differentiated cells. PLoS Pathog 9:e1003321
McBride, Alison A; Sakakibara, Nozomi; Stepp, Wesley H et al. (2012) Hitchhiking on host chromatin: how papillomaviruses persist. Biochim Biophys Acta 1819:820-5
Sakakibara, Nozomi; Mitra, Ruchira; McBride, Alison A (2011) The papillomavirus E1 helicase activates a cellular DNA damage response in viral replication foci. J Virol 85:8981-95

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