The energetics of specific binding of AT-hooks from HMGA1 to target DNA

A. Dragan, J. Liggins, Colyn Crane-Robinson, P. Privalov

Research output: Contribution to journalArticlepeer-review


The interaction of the second and third AT-hooks of HMGA1 (formerly HMGI/Y), which bind selectively in the minor groove of an AT-rich DNA sequence, was studied at different temperatures and ionic strengths by spectropolarimetry, spectrofluorimetry, isothermal titration calorimetry and differential scanning calorimetry. The data show that binding of the ten amino acid core element of the two AT-hooks, which penetrates deep into the minor groove, is entropically driven: both the entropy and enthalpy of association of the peptides to the target DNA are positive up to 50 8C. The seven amino acid extension of the core in the secondAT-hook,which extends out from the minor groove and loops over the phosphodiester backbone, adds a substantial negative enthalpic component into the binding of the 17 residue DBD2 peptide to DNA that corresponds in magnitude to the enthalpy of formation of two hydrogen bonds. The ionic strength dependence of the association constant allowed an estimation of the electrostatic component of binding and, by subtraction, the contribution of the nonelectrostatic component, which results from dehydration of the contacting surfaces and makes up almost 70% of the total energy of complex formation. The exceptionally large positive entropy and enthalpy of association of the core AT-hook peptides with target DNA suggest that the water, which is removed from the minor groove of DNA upon binding, is in a highly ordered state. Acetylation of the lysine residue in the second AT-hook, which corresponds to Lys65 ofHMGA1, has little effect on the DNA binding; so it appears that repression of the hIFNb gene, which follows this modification, is not a direct result of the abrogation of DNA binding.
Original languageEnglish
Pages (from-to)393-411
Number of pages19
JournalJournal of Molecular Biology
Issue number2
Publication statusPublished - 21 Mar 2003


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