Walkinshaw M D, Taylor P, Sturrock S S, Atanasiu C, Berge T, Henderson R M, Edwardson J M, Dryden D T F
Institute of Cell and Molecular Biology, The King's Buildings, University of Edinburgh, EH9 3JR, Edinburgh, United Kingdom.
Mol Cell. 2002 Jan;9(1):187-94. doi: 10.1016/s1097-2765(02)00435-5.
We have solved, by X-ray crystallography to a resolution of 1.8 A, the structure of a protein capable of mimicking approximately 20 base pairs of B-form DNA. This ocr protein, encoded by gene 0.3 of bacteriophage T7, mimics the size and shape of a bent DNA molecule and the arrangement of negative charges along the phosphate backbone of B-form DNA. We also demonstrate that ocr is an efficient inhibitor in vivo of all known families of the complex type I DNA restriction enzymes. Using atomic force microscopy, we have also observed that type I enzymes induce a bend in DNA of similar magnitude to the bend in the ocr molecule. This first structure of an antirestriction protein demonstrates the construction of structural mimetics of long segments of B-form DNA.
我们通过X射线晶体学解析出了一种能够模拟大约20个碱基对B型DNA的蛋白质结构,分辨率达到1.8埃。这种由噬菌体T7的0.3基因编码的ocr蛋白,模拟了弯曲DNA分子的大小和形状以及B型DNA磷酸主链上负电荷的排列。我们还证明,ocr在体内是所有已知复合型I类DNA限制酶家族的有效抑制剂。利用原子力显微镜,我们还观察到I类酶会使DNA产生与ocr分子中弯曲程度相似的弯曲。这种抗限制蛋白的首个结构展示了B型DNA长片段结构模拟物的构建。