Anasuya Chattopadhyay, Martin C. Schmidt, Saleem A. Khan
Department of Molecular Genetics and Biochemistry
University of Pittsburgh School of Medicine, Pittsburgh, PA 15261
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 3V 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 5V region of the viral LCR and 157 nucleotides from the
3V 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.
