Roychoudhury M, Sitlani A, Lapham J, Crothers D M
Department of Chemistry, P.O. Box 208107, Yale University, New Haven, CT 06520, USA.
Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13608-13. doi: 10.1073/pnas.250476297.
The method of DNA cyclization kinetics reveals special properties of the TATAAACGCC sequence motif found in DNA sequences that have high affinity for core histones. Replacement of 30 bp of generic DNA by three 10-bp repeats of the motif in small cyclization constructs increases cyclization rates by two orders of magnitude. We document a 13 degrees bend in the motif and characterize the direction of curvature. The bending force constant is smaller by nearly 2-fold and there is a 35% decrease in the twist modulus, relative to generic DNA. These features are the likely source of the high affinity for bending around core histones to form nucleosomes. Our results establish a protocol for determination of the ensemble-averaged global solution structure and mechanical properties of any approximately 10-bp DNA sequence element of interest, providing information complementary to that from NMR and crystallographic structural studies.
DNA环化动力学方法揭示了在对核心组蛋白具有高亲和力的DNA序列中发现的TATAAACGCC序列基序的特殊性质。在小环化构建体中,用该基序的三个10碱基对重复序列替换30碱基对的通用DNA,可使环化速率提高两个数量级。我们记录了该基序中13度的弯曲,并表征了曲率方向。相对于通用DNA,弯曲力常数小近2倍,扭转模量降低35%。这些特征可能是对围绕核心组蛋白弯曲以形成核小体具有高亲和力的来源。我们的结果建立了一种协议,用于确定任何感兴趣的约10碱基对DNA序列元件的整体平均全局溶液结构和力学性质,提供与NMR和晶体学结构研究互补的信息。