title: Determination of the DNA sequence recognized by the bHLH-zip domain of the N-Myc protein.
abstract: The DNA-binding domain of the murine N-Myc protein, comprising the basic helix-loop-helix-zipper (bHLH-zip) region was expressed as a fusion protein in E. coli. The affinity purified glutathione-S-transferase-N-Myc fusion protein (GST-N-MYC) was used to select the N-Myc specific DNA-recognition motif from a pool of random-sequence oligonucleotides. After seven rounds of binding-site selection, specifically enriched oligonucleotides were cloned and sequenced. Of 31 individual oligonucleotides whose sequences were determined, 30 contained a common DNA-motif, defining the hexameric consensus sequence CACGTG. We confirm by mutational analysis that binding of the N-Myc derived bHLH-zip domain to this motif is sequence-specific.
authors.affiliation: Institute for Genetics, University of Cologne, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Alex
authors.forename: O
authors.initials: O
authors.lastname: Sözeri
authors.forename: S
authors.initials: S
authors.lastname: Meyer
authors.forename: R
authors.initials: R
authors.lastname: Dildrop
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D016259:Genes, myc; D005982:Glutathione Transferase / Q000235:genetics; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
chemical_list: D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins; D004247:DNA; D005982:Glutathione Transferase