Abstract
The KAP-2 protein that binds to the H-box (CCTACC) element in the bean CHS15 chalcone synthase promoter was purified, and internal peptide sequence used to design primers leading to the cloning of KAP-2 from bean (Phaseolus vulgaris) and barrel medic (Medicago truncatula). KAP-2 shares sequence similarity to the large subunit of mammalian Ku autoantigen, a protein proposed to be involved in control of DNA recombination and transcription. KAP-2 sequences were present in genomic DNA from a range of legumes, and a related protein is found in Arabidopsis thaliana. Recombinant KAP-2 expressed in insect cells showed the same binding specificity for the CHS15 H-box as the protein purified from bean cell extracts. In vitro transcription assays confirmed that KAP-2 stimulates transcription from a promoter harboring the H-box cis element.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acyltransferases / genetics*
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Acyltransferases / metabolism
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Amino Acid Sequence
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Animals
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Antigens, Nuclear*
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Base Sequence
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Binding Sites / genetics
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Cell Line
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DNA Helicases*
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DNA, Complementary / chemistry
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DNA, Complementary / genetics
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DNA, Complementary / isolation & purification
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DNA, Plant / genetics
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DNA-Binding Proteins / genetics*
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Fabaceae / enzymology*
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Fabaceae / genetics
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Fabaceae / microbiology
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Gene Expression Regulation
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Humans
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Ku Autoantigen
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Medicago / genetics
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Medicago / microbiology
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Molecular Sequence Data
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Nuclear Proteins / genetics*
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Oligonucleotides / genetics
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Oligonucleotides / metabolism
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Phaseolus / enzymology
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Phaseolus / genetics
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Phaseolus / microbiology
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Plant Proteins / genetics*
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Plant Proteins / metabolism
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Promoter Regions, Genetic / genetics*
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Protein Binding
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Recombinant Fusion Proteins / metabolism
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Sequence Alignment
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Sequence Analysis, DNA
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Sequence Analysis, Protein
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Sequence Homology, Amino Acid
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Sinorhizobium meliloti / growth & development
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Spodoptera / cytology
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Spodoptera / genetics
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Transcription Factors / genetics*
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Transcription Factors / metabolism
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Transcription, Genetic / genetics
Substances
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Antigens, Nuclear
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DNA, Complementary
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DNA, Plant
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DNA-Binding Proteins
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Nuclear Proteins
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Oligonucleotides
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Plant Proteins
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Recombinant Fusion Proteins
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Transcription Factors
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Acyltransferases
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flavanone synthetase
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DNA Helicases
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XRCC5 protein, human
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Xrcc6 protein, human
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Ku Autoantigen
Associated data
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GENBANK/AF293343
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GENBANK/AF293344