Identification of a 450-bp region of human papillomavirus type 1 that promotes episomal replication in Saccharomyces cerevisiae.
Human papillomaviruses (HPVs) replicate as nuclear plasmids in infected cells. Since the DNA replication machinery is generally conserved between humans and Saccharomyces cerevisiae, we studied whether HPV-1 DNA can replicate in yeast. Plasmids containing a selectable marker (with or without a yeast centromere) and either the full-length HPV-1 genome or various regions of the viral long control region (LCR) and the 3' end of the L1 gene were introduced into S. cerevisiae and their ability to replicate episomally was investigated. Our results show that HPV-1 sequences promote episomal replication of plasmids although the yeast centromere is required for plasmid retention. We have mapped the autonomously replicating sequence activity of HPV-1 DNA to a 450 base-pair sequence (HPV-1 nt 6783-7232) that includes 293 nucleotides from the 5' region of the viral LCR and 157 nucleotides from the 3' end of the L1 gene. The HPV-1 ARS does not include the binding sites for the viral E1 and E2 proteins, and these proteins are dispensable for replication in S. cerevisiae.
Chattopadhyay
Anasuya
A
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Room East 1240 Biomedical Science Tower, Pittsburgh, PA 15261, USA.
Schmidt
Martin C
MC
Khan
Saleem A
SA
D020029:Base Pairing; D015139:Blotting, Southern; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics*; D016679:Genome, Viral; D027383:Papillomaviridae / Q000235:genetics / Q000502:physiology*; D010957:Plasmids; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000821:virology*; D014779:Virus Replication / Q000502:physiology*
D004279:DNA, Viral