Abstract
Computer modeling suggested that transcription factors with novel sequence specificities could be designed by combining known DNA binding domains. This structure-based strategy was tested by construction of a fusion protein, ZFHD1, that contained zinc fingers 1 and 2 from Zif268, a short polypeptide linker, and the homeodomain from Oct-1. The fusion protein bound optimally to a sequence containing adjacent homeodomain (TAATTA) and zinc finger (NGGGNG) subsites. When fused to an activation domain, ZFHD1 regulated promoter activity in vivo in a sequence-specific manner. Analysis of known protein-DNA complexes suggests that many other DNA binding proteins could be designed in a similar fashion.
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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Amino Acid Sequence
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Base Sequence
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Binding Sites
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Cloning, Molecular
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Computer Simulation
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Gene Expression Regulation
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Homeodomain Proteins / chemistry
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Host Cell Factor C1
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Models, Molecular
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Molecular Sequence Data
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Octamer Transcription Factor-1
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Promoter Regions, Genetic
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Protein Engineering
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / metabolism
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Transcription Factors / chemistry*
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Transfection
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Zinc Fingers*
Substances
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DNA-Binding Proteins
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Homeodomain Proteins
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Host Cell Factor C1
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Octamer Transcription Factor-1
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Recombinant Fusion Proteins
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Transcription Factors
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ZFHD1 protein, recombinant