Fields D S, He Y, Al-Uzri A Y, Stormo G D
Department of Molecular Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309-0347, USA.
J Mol Biol. 1997 Aug 15;271(2):178-94. doi: 10.1006/jmbi.1997.1171.
The Mnt protein of Salmonella phage P22 binds site-specifically to its operator. To better understand this binding we used dideoxy DNA sequencing in a quantitative manner to determine the relative binding constants, and hence the relative free energies, of wild-type Mnt protein to a substantial number of variants of its operator. These measurements were supported by experiments which used the SELEX procedure to generate a set of operators from an initially randomized population. In the Discussion we show that the present model of Mnt protein/operator binding, due to Sauer and co-workers, along with the assumption of an independent contribution of each position in the operator to the total binding, provides a reasonably accurate description of the system. We also discuss the use of information content as a measure of DNA-protein binding specificity with the Mnt protein/operator system serving as an example and show again that the assumption of independence supports the current view of this case of site-specific binding.
沙门氏菌噬菌体P22的Mnt蛋白以位点特异性方式与其操纵子结合。为了更好地理解这种结合,我们以定量方式使用双脱氧DNA测序来确定野生型Mnt蛋白与大量操纵子变体的相对结合常数,进而确定相对自由能。这些测量得到了实验的支持,这些实验使用SELEX程序从最初随机的群体中生成了一组操纵子。在讨论中,我们表明,由于绍尔及其同事提出的当前Mnt蛋白/操纵子结合模型,以及操纵子中每个位置对总结合的独立贡献的假设,为该系统提供了合理准确的描述。我们还以Mnt蛋白/操纵子系统为例讨论了使用信息含量作为DNA-蛋白质结合特异性的度量,并再次表明独立性假设支持了这种位点特异性结合情况的当前观点。