RecA-double stranded DNA complexes studied by atomic force microscopy.

K Umemura, J Komatsu, T Uchihashi, N Choi, S Ikawa, T Nishinaka, T Shibata, Y Nakayama, S Katsura, A Mizuno, H Tokumoto, M Ishikawa, R Kuroda
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Abstract

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.

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原子力显微镜研究reca -双链DNA复合物。
用原子力显微镜(AFM)研究了reca -双链(ds) DNA复合物。当在ATP γ S存在下制备复合物时,AFM观察到完全覆盖的RecA-dsDNA细丝。当RecA蛋白浓度较低时,可以发现不同长度的细丝。观察到的结构变化可以直接反映反应混合物中中间配合物的真实分布,因为混合物只是简单地沉积在云母表面进行AFM观察,没有特殊的固定或染色。碳纳米管(CNT)原子力显微镜尖端的使用可以高分辨率地揭示RecA-dsDNA细丝的周期性。我们的观察证明了AFM方法在RecA-dsDNA复合物结构研究中的潜力,特别是它们的中间态。
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