Pang D, Yoo S, Dynan W S, Jung M, Dritschilo A
Department of Radiation Medicine, Georgetown University Medical Center, Washington, DC 20007-2197, USA.
Cancer Res. 1997 Apr 15;57(8):1412-5.
The binding of the Ku protein to DNA was investigated using the atomic force microscope. Ku was found to bind predominantly to the ends of double-stranded DNA. Experiments with plasmid DNA revealed that Ku does not bind to circular plasmids but does bind to plasmids that have been linearized by treatment with ionizing radiation. The binding of Ku to poly(dG-dC) x poly(dG-dC) polynucleotides and to a 400-bp DNA EcoRI fragment resulted in a shift in the fragment size distribution to include longer fragments, with internally binding Ku. Furthermore, we observed images consistent with fragments joined together by Ku, showing an interaction with two ends of DNA. These observations suggest that Ku may play a role in physically orienting DNA for ligation by binding the ends of adjacent DNA molecules.
利用原子力显微镜研究了Ku蛋白与DNA的结合情况。发现Ku主要结合于双链DNA的末端。对质粒DNA进行的实验表明,Ku不与环状质粒结合,但确实能与经电离辐射处理而线性化的质粒结合。Ku与聚(dG-dC)×聚(dG-dC)多核苷酸以及400碱基对的DNA EcoRI片段的结合导致片段大小分布发生偏移,出现了更长的片段,这是由于Ku在内侧结合。此外,我们观察到的图像与通过Ku连接在一起的片段相符,显示出与DNA两端的相互作用。这些观察结果表明,Ku可能通过结合相邻DNA分子的末端,在物理上使DNA定向以便进行连接方面发挥作用。