Journal Article

The DNA Bend Angle and Binding Affinity of an HMG Box Increased by the Presence of Short Terminal Arms

Mirna Lnenicek-Allen, Christopher M. Read and Colyn Crane-Robinson

in Nucleic Acids Research

Volume 24, issue 6, pages 1047-1051
Published in print March 1996 | ISSN: 0305-1048
Published online March 1996 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/24.6.1047
The DNA Bend Angle and Binding Affinity of an HMG Box Increased by the Presence of Short Terminal Arms

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The HMG box of human LEF-1 (hLEF-1, formerly TCF1α) 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 a-chain enhancer. In the bending assay, whereas the parent box and that with the N-terminal extension bent the DNA by 57/58°, the box extended at the C-terminus bent the DNA by 77/78°, 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.

Journal Article.  2864 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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