Starr D B, Hoopes B C, Hawley D K
Department of Chemistry, University of Oregon, Eugene 97403, USA.
J Mol Biol. 1995 Jul 21;250(4):434-46. doi: 10.1006/jmbi.1995.0388.
We have used gel electrophoretic methods to analyze the extent, location and direction of the DNA bend induced by the TATA binding protein (TBP) upon binding to a consensus TATA box sequence. Our observations were consistent with the proposed models for the X-ray crystal structure of the TBP-TATA box complex. We have also measured the magnitude and direction of the bend induced by TBP upon binding a number of variant TATA box sequences for which we have measured TBP binding affinity. We found that the extent to which the DNA was bent in the complex differed among the various sequences and was correlated with the stability of the complex; that is, the greater the stability of the complex, the more the DNA appeared to be bent by TBP. This study provides the first evidence that the structure of the TBP-DNA complex may vary with different DNA sequences. In addition, we propose, based on our findings, that the energetics of bending contribute significantly to the overall binding affinity of TBP for different sequences.
我们运用凝胶电泳方法,分析了TATA结合蛋白(TBP)与共有TATA框序列结合时所诱导的DNA弯曲的程度、位置及方向。我们的观察结果与TBP-TATA框复合物的X射线晶体结构的推测模型相符。我们还测定了TBP与多个变异TATA框序列结合时所诱导的弯曲的幅度和方向,对于这些变异序列,我们已测定了TBP的结合亲和力。我们发现,在复合物中DNA的弯曲程度在不同序列间存在差异,且与复合物的稳定性相关;也就是说,复合物的稳定性越高,TBP使DNA弯曲的程度似乎就越大。这项研究首次证明了TBP-DNA复合物的结构可能因不同的DNA序列而有所不同。此外,基于我们的研究结果,我们提出,弯曲的能量学对TBP与不同序列的整体结合亲和力有显著贡献。