van Beest M, Dooijes D, van De Wetering M, Kjaerulff S, Bonvin A, Nielsen O, Clevers H
Department of Immunology, University Medical Center Utrecht, Heidelberglaan 100 Rm F03.821, 3584 CX Utrecht, The Netherlands.
J Biol Chem. 2000 Sep 1;275(35):27266-73. doi: 10.1074/jbc.M004102200.
Sequence-specific high mobility group (HMG) box factors bind and bend DNA via interactions in the minor groove. Three-dimensional NMR analyses have provided the structural basis for this interaction. The cognate HMG domain DNA motif is generally believed to span 6-8 bases. However, alignment of promoter elements controlled by the yeast genes ste11 and Rox1 has indicated strict conservation of a larger DNA motif. By site selection, we identify a highly specific 12-base pair motif for Ste11, AGAACAAAGAAA. Similarly, we show that Tcf1, MatMc, and Sox4 bind unique, highly specific DNA motifs of 12, 12, and 10 base pairs, respectively. Footprinting with a deletion mutant of Ste11 reveals a novel interaction between the 3' base pairs of the extended DNA motif and amino acids C-terminal to the HMG domain. The sequence-specific interaction of Ste11 with these 3' base pairs contributes significantly to binding and bending of the DNA motif.
序列特异性高迁移率族（HMG）盒因子通过在小沟中的相互作用与DNA结合并使其弯曲。三维核磁共振分析为这种相互作用提供了结构基础。同源HMG结构域DNA基序一般认为跨越6至8个碱基。然而，由酵母基因ste11和Rox1控制的启动子元件比对表明，存在一个更大的DNA基序的严格保守性。通过位点选择，我们确定了Ste11的一个高度特异性的12碱基对基序，即AGAACAAAGAAA。同样，我们表明Tcf1、MatMc和Sox4分别结合独特的、高度特异性的12、12和10碱基对的DNA基序。用Ste11的缺失突变体进行足迹分析揭示了延伸DNA基序的3'端碱基对与HMG结构域C端氨基酸之间的一种新的相互作用。Ste11与这些3'端碱基对的序列特异性相互作用对DNA基序的结合和弯曲有显著贡献。
