Abstract
By computer search, we identified one potential NF-kappaB binding site in the HPV16 long control region (LCR) at position 7554-7563 having two mismatches in comparison to the consensus NF-kappaB binding site of the Igkappa L promoter. Bandshift experiments with nuclear extracts from HeLa cells or purified glutathione S-transferase-p65 fusion protein clearly demonstrated that NF-kappaB is able to bind to this region of the LCR. However, in comparison to NF-kappaB binding on a consensus probe, the affinity of NF-kappaB for this site is about 250-fold reduced. When mutations were introduced into this NF-kappaB binding site, the activity of the LCR was increased, strongly suggesting that NF-kappaB was acting as a transcriptional repressor in the context of the HPV16 LCR. In addition, overexpression of NF-kappaB p65 repressed the activity of the HPV16 LCR, strengthening this conclusion.
Copyright 2000 Academic Press.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Base Sequence
-
Binding Sites
-
CCAAT-Enhancer-Binding Proteins
-
Consensus Sequence / genetics
-
DNA Probes / genetics
-
DNA Probes / metabolism
-
DNA, Viral / genetics
-
DNA, Viral / metabolism*
-
DNA-Binding Proteins / metabolism
-
Gene Expression Regulation, Viral / genetics*
-
HeLa Cells
-
Humans
-
Mutation / genetics
-
NF-kappa B / genetics
-
NF-kappa B / metabolism*
-
NFI Transcription Factors
-
Nuclear Proteins / metabolism
-
Oligodeoxyribonucleotides / genetics
-
Oligodeoxyribonucleotides / metabolism
-
Papillomaviridae / genetics*
-
Promoter Regions, Genetic / genetics*
-
Protein Binding
-
Recombinant Fusion Proteins / metabolism
-
Repressor Proteins / genetics
-
Repressor Proteins / metabolism
-
Response Elements / genetics*
-
Thermodynamics
-
Transcription Factor RelA
-
Transcription Factors / metabolism
-
Transfection
-
Tumor Cells, Cultured
Substances
-
CCAAT-Enhancer-Binding Proteins
-
DNA Probes
-
DNA, Viral
-
DNA-Binding Proteins
-
NF-kappa B
-
NFI Transcription Factors
-
Nuclear Proteins
-
Oligodeoxyribonucleotides
-
Recombinant Fusion Proteins
-
Repressor Proteins
-
Transcription Factor RelA
-
Transcription Factors
-
transcription factor nuclear factor 1