title: The DNA sequence specificity of HMG boxes lies in the minor wing of the structure.
abstract: To establish the basis of sequence-specific DNA recognition by HMG boxes we separately transferred the minor and major wings from the sequence-specific HMG box of TCF1 alpha into their equivalent position in the non-sequence-specific box 2 of HMG1. Thus chimera THT1 contains the minor wing (of 11 N-terminal and 25 C-terminal residues) from the HMG box of TCF1 alpha and the major wing (the 45 residue central section) from HMG1 box 2, whilst the situation is reversed in chimera HTH1. The structural integrity of the two chimeric proteins was established by CD, NMR and their binding to four-way junction DNA. Gel retardation and circular permutation assays showed that only chimera THT1, containing the TCF1 alpha minor wing, formed a sequence-specific complex and bent the DNA. The bend angle was estimated to be 59 degrees for chimera THT1 and 52 degrees for the HMG box of TCF1 alpha. Our results, in combination with mutagenesis and other data, suggests a model for the DNA binding of HMG boxes in which the N-terminal residues and part of helix 1 contact the minor groove on the outside of a bent DNA duplex.
authors.affiliation: Biophysics Laboratories, University of Portsmouth, UK.
authors.forename: C M
authors.initials: CM
authors.lastname: Read
authors.forename: P D
authors.initials: PD
authors.lastname: Cary
authors.forename: N S
authors.initials: NS
authors.lastname: Preston
authors.forename: M
authors.initials: M
authors.lastname: Lnenicek-Allen
authors.forename: C
authors.initials: C
authors.lastname: Crane-Robinson
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid
chemical_list: D006609:High Mobility Group Proteins; D004247:DNA