Abstract
The human papillomavirus (HPV)-16 oncogenes, E6 and E7, are transcribed preferentially in keratinocytes and cervical carcinoma cells due to a 5' enhancer. An abundant peptide binding to a 37 nt enhancer element was purified from human keratinocytes by sequence-specific DNA chromatography. This protein was identified as transcriptional enhancer factor (TEF)-1 by complex mobility, binding to wild-type and mutant SV40 and HPV-16 enhansons and antigenic reactivity with two anti-TEF-1 antibodies. TEF-1 is cell-specific, but its transactivation also depends on a limiting, cell-specific TEF-1 'co-activator'. We show that both TEF-1 and the TEF-1 co-activator are active in human keratinocytes and essential for HPV-16 transcription. TEF-1 binding in vivo was necessary for HPV-16 P97 promoter activity. Excess TEF-1 and chimeric GAL4-TEF-1 specifically inhibited the P97 promoter by 'squelching', indicating that HPV-16 transcription also requires a limiting TEF-1 co-activator. TEF-1 and the TEF-1 co-activator functions mirrored HPV-16 transcription by their presence in keratinocytes and cervical carcinoma cells and their absence from lymphoid B-cells, but also functioned in liver cells where the HPV-16 promoter is inactive. TEF-1 and its associated co-activator are thus part of a complex mechanism which determines the restricted cell range of the HPV-16 E6 and E7 oncogene promoter.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Antibodies
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Base Sequence
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Cell Line
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Cell Nucleus / physiology
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Chloramphenicol O-Acetyltransferase / genetics
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Chloramphenicol O-Acetyltransferase / metabolism
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Chromatography, Affinity
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / isolation & purification
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DNA-Binding Proteins / metabolism*
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Enhancer Elements, Genetic
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Female
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Gene Expression Regulation, Viral*
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Humans
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Keratinocytes / physiology*
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Nuclear Proteins*
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Oligodeoxyribonucleotides
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Oncogene Proteins, Viral / genetics*
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Oncogenes*
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Papillomaviridae / genetics*
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Papillomavirus E7 Proteins
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Promoter Regions, Genetic
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Recombinant Fusion Proteins / metabolism
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Repressor Proteins*
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TEA Domain Transcription Factors
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Transcription Factors / genetics
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Transcription Factors / isolation & purification
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Transcription Factors / metabolism*
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Transcription, Genetic*
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Transcriptional Activation*
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Transfection
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Uterine Cervical Neoplasms / genetics*
Substances
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Antibodies
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DNA-Binding Proteins
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E6 protein, Human papillomavirus type 16
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Nuclear Proteins
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Oligodeoxyribonucleotides
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Oncogene Proteins, Viral
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Papillomavirus E7 Proteins
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Recombinant Fusion Proteins
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Repressor Proteins
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TEA Domain Transcription Factors
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TEAD1 protein, human
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Transcription Factors
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oncogene protein E7, Human papillomavirus type 16
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Chloramphenicol O-Acetyltransferase
Associated data
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GENBANK/X64700
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GENBANK/X65365
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GENBANK/X65366
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GENBANK/X65367
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GENBANK/X65368
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GENBANK/X66269
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GENBANK/X67723
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GENBANK/X67724
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GENBANK/X67725
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GENBANK/X67726