Abstract
Over one thousand mutations have been identified to date in the cystic fibrosis transmembrane conductance regulator gene ( CFTR); however, about 5-10% of mutations remain undefined. It is likely that some of these undefined mutations occur within regulatory elements for the CFTR gene. Tissue-specific regulatory elements for CFTR are located outside the basal promoter region and may be associated with DNase I hypersensitive sites (DHS). We previously described a DHS at +15.6 kb 3' to the CFTR gene, which showed tissue specificity in vivo and was evaluated as a candidate regulatory element for CFTR. Polymorphisms in regulatory elements may have a significant effect on their activity and hence on the levels of gene expression. Two C-->T polymorphisms were identified within the +15.6 kb region that occurred on both cystic fibrosis (CF) and non-CF alleles. Both of the polymorphisms altered DNA-protein binding, as shown by electrophoretic mobility shift assays (EMSA). These changes in transcription factor binding at a putative regulatory region could influence CFTR gene expression.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
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Binding Sites
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Binding, Competitive
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COUP Transcription Factors
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Case-Control Studies
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Cells, Cultured
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Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
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DNA Footprinting
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DNA Primers / chemistry
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DNA-Binding Proteins / metabolism
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Deoxyribonuclease I
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Electrophoretic Mobility Shift Assay
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Gene Expression Regulation
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Hepatocyte Nuclear Factor 4
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Humans
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Molecular Sequence Data
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Mutation
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Oligonucleotides / metabolism
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Phosphoproteins / metabolism
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Polymerase Chain Reaction
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Polymorphism, Genetic*
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Polymorphism, Single-Stranded Conformational
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Promoter Regions, Genetic / genetics*
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Protein Binding
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Receptors, Steroid*
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Regulatory Sequences, Nucleic Acid
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Transcription Factors / metabolism
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Transcription, Genetic
Substances
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
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CFTR protein, human
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COUP Transcription Factors
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DNA Primers
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DNA-Binding Proteins
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Hepatocyte Nuclear Factor 4
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MLX protein, human
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Oligonucleotides
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Phosphoproteins
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Receptors, Steroid
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Transcription Factors
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Cystic Fibrosis Transmembrane Conductance Regulator
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Deoxyribonuclease I