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J Virol, June 1998, p. 4911-4917, Vol. 72, No. 6
Department of Biochemistry and Molecular
Genetics, University of Alabama at Birmingham, Birmingham, Alabama
35294
Received 29 August 1997/Accepted 5 March 1998
We have established cell-free replication for the human
papillomavirus type 18 (HPV-18) origin of replication
(ori)-containing DNA by using purified HPV-18 E1 and E2
gene products expressed as fusion proteins in Escherichia
coli. The transcription factor YY1 has been shown to regulate RNA
transcription by binding to a sequence overlapping the putative E1
protein binding site in the HPV-18 ori. We show that
exogenously added YY1 fusion protein inhibited HPV-18 ori
replication. Cotransfection of YY1 expression vectors also inhibited
transient replication in 293 cells. However, inhibition did not appear
to be mediated by binding to its cognate site in the ori as
YY1 also inhibited the replication of the HPV-11 ori, which
does not have a known or suspected YY1 binding site. Moreover,
inhibition was not alleviated by the inclusion of YY1 binding
oligonucleotides in the replication reaction mixtures. Rather, we
demonstrated a direct interaction between purified fusion E2 protein
and fusion YY1 protein by the pull-down assay and a partial restoration
of replication activity by an elevated E2 protein concentration. These
results suggest that YY1 can inhibit HPV ori replication by
interfering with E2 protein functions.
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Transcription Factor YY1 Represses Cell-Free
Replication from Human Papillomavirus Origins
*
Corresponding author. Mailing address: Department of
Biochemistry and Molecular Genetics, University of Alabama at
Birmingham, 1918 University Blvd., Birmingham, AL 35294-0005. Phone:
(205) 975-8300. Fax: (205) 975-6075. E-mail: LTChow{at}uab.edu.
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