Abstract
Polypyrimidine tract binding (PTB) protein is a cellular factor whose function is unknown. Various RNA or single-stranded DNA sequences have been shown to interact with PTB. In this paper, using laser UV crosslinking and electrophoretic mobility shift assays to probe DNA-protein interactions, we demonstrate that PTB binding at a single-stranded DNA target is highly sequence-specific. We provide data showing that PTB interacts with the top strand of the adenovirus major late promoter transcriptional initiator, a sequence rich in pyrimidine residues. We also demonstrate that PTB is organised into at least two different binding domains.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenoviridae / genetics
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Base Sequence
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Binding, Competitive
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Cross-Linking Reagents
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DNA, Single-Stranded / chemistry*
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DNA, Single-Stranded / metabolism
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DNA-Binding Proteins / chemistry*
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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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Escherichia coli / genetics
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Kinetics
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Molecular Sequence Data
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Molecular Weight
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Oligodeoxyribonucleotides / chemistry
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Plasmids / chemistry
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Polypyrimidine Tract-Binding Protein
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Promoter Regions, Genetic
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RNA-Binding Proteins / chemistry*
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RNA-Binding Proteins / genetics
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RNA-Binding Proteins / isolation & purification
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RNA-Binding Proteins / metabolism
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Ultraviolet Rays
Substances
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Cross-Linking Reagents
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DNA, Single-Stranded
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DNA-Binding Proteins
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Oligodeoxyribonucleotides
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RNA-Binding Proteins
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Recombinant Fusion Proteins
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Polypyrimidine Tract-Binding Protein