Using a Human Papillomavirus Model to Study DNA Replication and Repair of Wild Type and Damaged DNA Templates in Mammalian Cells

Human papillomaviruses have 8kbp DNA episomal genomes that replicate autonomously from host DNA. During initial infection, the virus increases its copy number to 20-50 copies per cell, causing torsional stress on the replicating DNA. This activates the DNA damage response (DDR) and HPV replicates it...

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Bibliographic Details
Published in:International journal of molecular sciences 2020-10, Vol.21 (20), p.7564
Main Authors: Das, Dipon, Bristol, Molly L, Pichierri, Pietro, Morgan, Iain M
Format: Article
Language:English
Subjects:
Alphapapillomavirus - genetics
Alphapapillomavirus - metabolism
Animals
Bacteria
Binding sites
Cancer
Cell cycle
Cervical cancer
Copy number
Deoxyribonucleic acid
DNA
DNA biosynthesis
DNA Damage
DNA helicase
DNA methylation
DNA Repair
DNA Replication
E1 and E2
Genetic Engineering - methods
Genomes
Head & neck cancer
Homologous recombination
Homology
Human papillomavirus
human papillomaviruses
Humans
Infections
LacZ gene
Mammalian cells
Mammals
model system
Molecular weight
Papillomaviridae
Plasmids
Proteins
Repair
Replication
replication and repair
Replication origins
Review
Torsional stress
Transfection
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Summary:Human papillomaviruses have 8kbp DNA episomal genomes that replicate autonomously from host DNA. During initial infection, the virus increases its copy number to 20-50 copies per cell, causing torsional stress on the replicating DNA. This activates the DNA damage response (DDR) and HPV replicates its genome, at least in part, using homologous recombination. An active DDR is on throughout the HPV life cycle. Two viral proteins are required for replication of the viral genome; E2 binds to 12bp palindromic sequences around the A/T rich origin of replication and recruits the viral helicase E1 via a protein-protein interaction. E1 forms a di-hexameric complex that replicates the viral genome in association with host factors. Transient replication assays following transfection with E1-E2 expression plasmids, along with an origin containing plasmid, allow monitoring of E1-E2 replication activity. Incorporating a bacterial lacZ gene into the origin plasmid allows for the determination of replication fidelity. Here we describe how we exploited this system to investigate replication and repair in mammalian cells, including using damaged DNA templates. We propose that this system has the potential to enhance the understanding of cellular components involved in DNA replication and repair.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms21207564