Hogan M, Dattagupta N, Crothers D M
Proc Natl Acad Sci U S A. 1978 Jan;75(1):195-9. doi: 10.1073/pnas.75.1.195.
We report transient electric dichroism studies on monodisperse rod-like DNA molecules. By using restriction fragments and DNAs of known length, it is shown that the orientation time is accurately predicted by the theoretically calculated rotational diffusion coefficient. The field dependence of the steady-state dichroism values is not consistent with the induced electric dipole orientation mechanism, and the time dependence is not consistent with the presence of a permanent dipole moment. In order to explain the dependence of the dichroism on the electric field, the ionic strength of the medium, and the length of the macromolecule, we propose a new model in which anisotropic ion flow produces an asymmetric ion atmosphere around the polyelectrolyte, resulting in an orienting torque. From the limiting dichroism at high field, we estimate that the DNA bases are inclined at an angle of 73 degrees or less relative to the helix axis, in good agreement with the revised model of B-form DNA suggested by Levitt, in which the base pairs have a propeller-like twist. Our results establish transient electric dichroism measurements as a technique well suited for study of alterations in the length and base pair inclination of rod-like DNA molecules.
我们报道了对单分散棒状DNA分子的瞬态电二色性研究。通过使用限制性片段和已知长度的DNA,结果表明,取向时间可由理论计算的旋转扩散系数准确预测。稳态二色性值的场依赖性与感应电偶极取向机制不一致,且时间依赖性与永久偶极矩的存在不一致。为了解释二色性对电场、介质离子强度和大分子长度的依赖性,我们提出了一个新模型,其中各向异性离子流在聚电解质周围产生不对称离子氛围,从而产生取向转矩。从高场下的极限二色性,我们估计DNA碱基相对于螺旋轴倾斜角度为73度或更小,这与Levitt提出的B型DNA修正模型非常吻合,在该模型中碱基对具有螺旋桨状扭曲。我们的结果表明,瞬态电二色性测量是一种非常适合研究棒状DNA分子长度和碱基对倾斜度变化的技术。