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Nucleic Acids Res ; 26(20): 4714-20, 1998 Oct 15.
Article in English | MEDLINE | ID: mdl-9753741

ABSTRACT

The maize ZmHox2a gene encodes two homeodomains which originated by a 699 bp duplication within an ancestral precursor. The sequences of the two ZmHox2a homeodomains are highly diverged in the N-terminal arm, while residues in the helical part have mostly been conserved. We show here that both ZmHox2a homeodomains are functional DNA-binding motifs but exhibit different target site specificities. CASTing experiments reveal a TCCT motif recognized by HD1 but a GATC tetranucleotide as the recognition sequence of HD2. Mutation of the central nucleotides in both tetranucleotide core motifs abolishes DNA binding. A domain swap experiment indicates that target site specificity is achieved in a combinatorial manner by the contributions of the diverged N-terminal arms together with the slightly different recognition helices. Computer modelling suggests that K47 and H54 in the recognition helices preferentially contact the bases at the 3'-terminus of the tetranucleotide target sequences.


Subject(s)
DNA-Binding Proteins/metabolism , Gene Duplication , Genes, Homeobox , Homeodomain Proteins/metabolism , Plant Proteins/metabolism , Zea mays/genetics , Amino Acid Sequence , Base Sequence , Binding Sites , Binding, Competitive , Computer Simulation , Consensus Sequence , DNA-Binding Proteins/genetics , Evolution, Molecular , Genes, Plant , Homeodomain Proteins/genetics , Molecular Sequence Data , Oligodeoxyribonucleotides/biosynthesis , Oligodeoxyribonucleotides/metabolism , Plant Proteins/genetics , Polymerase Chain Reaction , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/isolation & purification , Substrate Specificity , Zea mays/metabolism
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