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Title:
Non-consensus GLI binding sites in Hedgehog target gene regulation | BMC Molecular Biology
Description:
Background The GLI transcription factors, mediators of the hedgehog signal bind with high affinity to the consensus sequence GACCACCCA. The affinity of variant single substitutions in GLI binding sites has been measured systematically, but the affinities of the variant binding sites appears low compared to the frequency of occurrence of variant sites in known GLI target gene promoters. Results We quantified transcriptional activation by GLI using PTCH1 promoter based luciferase reporters containing all single substitutions of the GLI consensus binding site. As expected variants with very low affinity did not activate the reporter. Many lower affinity binding sequences are, however, functional in the presence of moderate GLI concentration. Using two natural non-consensus GLI site promoters we showed that substitution of the variant sequences by consensus leads to comparable activity. Conclusions Variant GLI binding sites with relatively low affinity can within natural promoters lead to strong transcriptional activation. This may facilitate the identification of additional direct GLI target genes.
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Keywords {🔍}
gli, binding, consensus, site, sites, activation, article, pubmed, reporter, sequence, promoter, luciferase, figure, google, scholar, gene, affinity, cas, ptch, variant, expression, target, transcription, construct, transcriptional, functional, activity, single, position, gliact, promoters, results, variants, genes, shown, constructs, human, cells, hedgehog, ptchvar, ptchwt, central, factors, sequences, control, wild, type, data, nonconsensus, substitutions,
Topics {✒️}
thomas eichberger & anna-maria frischauf sandra laner-plamberger open access article protein-dna binding interactions designer zinc-finger proteins finger gli-dna complex article download pdf pen/strep100x stock solution anna-maria frischauf introducing site-specific mutations hedgehog-responsive enhancers linked gli-dna binding domain transcription-factor binding affinity polyclonal goat-anti-gli2 laner-plamberger hedgehog/gli signal transduction full size image hh pathway activation author information authors molecular biology reporter gene transcription gli2-specific transcriptional activation reporter gene assays firefly luciferase gene regulate target genes affinity profile luciferase reporter system luciferase reporter assays transcription factor binding luciferase reporter assay luciferase reporter constructs dna binding domain gli binding site demonstrates specific binding view gli gene encodes hedgehog receptor ptch1 privacy choices/manage cookies ptch1_var luciferase reporter variant binding site binding site variant additional transcription factors wild type promoters gli binding sites dna binding sites authors’ original file gli transcription factors site directed mutagenesis selected binding sequences specific promoters attention
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headline:Non-consensus GLI binding sites in Hedgehog target gene regulation
description:The GLI transcription factors, mediators of the hedgehog signal bind with high affinity to the consensus sequence GACCACCCA. The affinity of variant single substitutions in GLI binding sites has been measured systematically, but the affinities of the variant binding sites appears low compared to the frequency of occurrence of variant sites in known GLI target gene promoters. We quantified transcriptional activation by GLI using PTCH1 promoter based luciferase reporters containing all single substitutions of the GLI consensus binding site. As expected variants with very low affinity did not activate the reporter. Many lower affinity binding sequences are, however, functional in the presence of moderate GLI concentration. Using two natural non-consensus GLI site promoters we showed that substitution of the variant sequences by consensus leads to comparable activity. Variant GLI binding sites with relatively low affinity can within natural promoters lead to strong transcriptional activation. This may facilitate the identification of additional direct GLI target genes.
datePublished:2010-01-13T00:00:00Z
dateModified:2010-01-13T00:00:00Z
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Luciferase Reporter
HaCaT Cell
Consensus Site
Luciferase Reporter Construct
Consensus Binding Site
Biochemistry
general
Nucleic Acid Chemistry
Cell Biology
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headline:Non-consensus GLI binding sites in Hedgehog target gene regulation
description:The GLI transcription factors, mediators of the hedgehog signal bind with high affinity to the consensus sequence GACCACCCA. The affinity of variant single substitutions in GLI binding sites has been measured systematically, but the affinities of the variant binding sites appears low compared to the frequency of occurrence of variant sites in known GLI target gene promoters. We quantified transcriptional activation by GLI using PTCH1 promoter based luciferase reporters containing all single substitutions of the GLI consensus binding site. As expected variants with very low affinity did not activate the reporter. Many lower affinity binding sequences are, however, functional in the presence of moderate GLI concentration. Using two natural non-consensus GLI site promoters we showed that substitution of the variant sequences by consensus leads to comparable activity. Variant GLI binding sites with relatively low affinity can within natural promoters lead to strong transcriptional activation. This may facilitate the identification of additional direct GLI target genes.
datePublished:2010-01-13T00:00:00Z
dateModified:2010-01-13T00:00:00Z
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HaCaT Cell
Consensus Site
Luciferase Reporter Construct
Consensus Binding Site
Biochemistry
general
Nucleic Acid Chemistry
Cell Biology
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