Umemura K, Komatsu J, Uchihashi T, Choi N, Ikawa S, Nishinaka T, Shibata T, Nakayama Y, Katsura S, Mizuno A, Tokumoto H, Ishikawa M, Kuroda R
Joint Research Center for Atom Technology, 1-1-4 Higashi, Tsukuba, Ibaraki 305-0046, Japan.
Nucleic Acids Symp Ser. 2000(44):213-4. doi: 10.1093/nass/44.1.213.
RecA-double stranded (ds) DNA complexes have been studied by atomic force microscopy (AFM). When the complexes were prepared in the presence of ATP gamma S, fully covered RecA-dsDNA filaments were observed by AFM. When the concentration of RecA proteins was lower, various lengths of filaments were found. The variation of the observed structures may directly reflect the real distribution of the intermediate complexes in the reaction mixture, as the mixture was simply deposited on a mica surface for AFM observation without special fixation or staining. The use of a carbon nanotube (CNT) AFM tip enabled high resolution to reveal the periodicity of RecA-dsDNA filaments. Our observations demonstrated the potential of the AFM method for the structural studies of the RecA-dsDNA complexes, especially their intermediate states.
已经通过原子力显微镜(AFM)对RecA双链(ds)DNA复合物进行了研究。当在ATPγS存在的情况下制备复合物时,通过AFM观察到完全覆盖的RecA-dsDNA细丝。当RecA蛋白的浓度较低时,发现了各种长度的细丝。观察到的结构变化可能直接反映了反应混合物中中间复合物的真实分布,因为该混合物只是简单地沉积在云母表面上用于AFM观察,而没有进行特殊固定或染色。使用碳纳米管(CNT)AFM探针能够实现高分辨率,以揭示RecA-dsDNA细丝的周期性。我们的观察结果证明了AFM方法在RecA-dsDNA复合物结构研究中的潜力,特别是它们的中间状态。