title: The DNA bend angle and binding affinity of an HMG box increased by the presence of short terminal arms.
abstract: The HMG box of human LEF-1 (hLEF-1, formerly TCF1alpha) has been expressed in four forms: a parent box of 81 amino acids and constructs having either a 10 amino acid C-terminal extension, a 9 amino acid N-terminal extension, or both. These four species have been compared for DNA binding and bending ability using a 28 bp recognition sequence from the TCR alpha-chain enhancer. In the bending assay, whereas the parent box and that with the N-terminal extension bent the DNA by 57/58 degrees, the box extended at the C-terminus bent the DNA by 77/78 degrees, irrespective of the presence or absence of the N-terminal extension. A 6- fold increase in DNA affinity also resulted from addition of both terminal extensions. These observations redefine the functional boundaries of the HMG box. The structure of a mouse LEF-1/DNA complex recently published [Love et al. (1995) Nature 376, 791-795] implies that the higher DNA affinity and in particular the increased bend angle observed are consequences, at least in part, of the C-terminal extension spanning the major groove on the inside of the DNA bend.
authors.affiliation: Biophysics Laboratories, University of Portsmouth, United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Lnenicek-Allen
authors.forename: C M
authors.initials: CM
authors.lastname: Read
authors.forename: C
authors.initials: C
authors.lastname: Crane-Robinson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D051821:Lymphoid Enhancer-Binding Factor 1; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; C497573:LEF1 protein, human; C497574:Lef1 protein, mouse; D051821:Lymphoid Enhancer-Binding Factor 1; D014157:Transcription Factors; D004247:DNA