Spronk C A, Folkers G E, Noordman A M, Wechselberger R, van den Brink N, Boelens R, Kaptein R
Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
EMBO J. 1999 Nov 15;18(22):6472-80. doi: 10.1093/emboj/18.22.6472.
The hinge-region of the lac repressor plays an important role in the models for induction and DNA looping in the lac operon. When lac repressor is bound to a tight-binding symmetric operator, this region forms an alpha-helix that induces bending of the operator. The presence of the hinge-helices is questioned by previous data that suggest that the repressor does not bend the wild-type operator. We show that in the wild-type complex the hinge-helices are formed and the DNA is bent, similar to the symmetric complex. Furthermore, our data show differences in the binding of the DNA binding domains to the half-sites of the wild-type operator and reveal the role of the central base-pair of the wild-type operator in the repressor-operator interaction. The differences in binding to the operator half-sites are incorporated into a model that explains the relative affinities of the repressor for various lac operator sequences that contain left and right half-sites with different spacer lengths.
乳糖阻遏蛋白的铰链区在乳糖操纵子的诱导和DNA环化模型中起着重要作用。当乳糖阻遏蛋白与紧密结合的对称操纵基因结合时,该区域形成一个α螺旋,诱导操纵基因弯曲。先前的数据对铰链螺旋的存在提出了质疑,这些数据表明阻遏蛋白不会使野生型操纵基因弯曲。我们表明,在野生型复合物中,铰链螺旋形成且DNA弯曲,类似于对称复合物。此外,我们的数据显示了DNA结合结构域与野生型操纵基因半位点结合的差异,并揭示了野生型操纵基因中心碱基对在阻遏蛋白-操纵基因相互作用中的作用。与操纵基因半位点结合的差异被纳入一个模型,该模型解释了阻遏蛋白对各种含有不同间隔长度的左右半位点的乳糖操纵基因序列的相对亲和力。