Abstract
The transcription factor Gli3 is acting mainly as a transcriptional repressor in the Sonic hedgehog signal transduction pathway. Gli3 contains a repressor domain in its N-terminus from residue G106 to E236. In this study we have characterized the intracellular structure of the Gli3 repressor domain using a combined bioinformatics and experimental approach. According to our findings the Gli3 repressor domain while being intrinsically disordered contains predicted anchor sites for partner interactions. The obvious interaction partners to test were Ski and DNA; however, with both of these the structure of Gli3 repressor domain remained disordered. To locate residues important for the repressor function we mutated several residues within the Gli3 repressor domain. Two of these, H141A and H157N, targeting predicted helical regions, significantly decreased transcriptional repression and thus identify important functional parts of the domain.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Cell Line
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DNA-Binding Proteins / metabolism
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Humans
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Intrinsically Disordered Proteins / chemistry
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Intrinsically Disordered Proteins / genetics
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Intrinsically Disordered Proteins / metabolism*
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Kruppel-Like Transcription Factors / chemistry
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Kruppel-Like Transcription Factors / genetics
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Kruppel-Like Transcription Factors / metabolism*
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Mutation
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Nerve Tissue Proteins / chemistry
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism*
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Nuclear Magnetic Resonance, Biomolecular
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Protein Binding
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Protein Interaction Domains and Motifs*
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Proto-Oncogene Proteins / metabolism
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Recombinant Fusion Proteins
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Repressor Proteins / chemistry
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Repressor Proteins / genetics
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Repressor Proteins / metabolism*
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Zinc Finger Protein Gli3
Substances
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DNA-Binding Proteins
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GLI3 protein, human
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Intrinsically Disordered Proteins
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Kruppel-Like Transcription Factors
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Nerve Tissue Proteins
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Proto-Oncogene Proteins
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Recombinant Fusion Proteins
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Repressor Proteins
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Zinc Finger Protein Gli3
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SKI protein, human