Hagerman P J
Nature. 1986;321(6068):449-50. doi: 10.1038/321449a0.
DNAs from both prokaryotic and eukaryotic organisms have yielded restriction fragments which manifest markedly anomalous electrophoretic behaviour (reduced mobility) when run on polyacrylamide gels. We have shown previously that the abnormal electrophoretic behaviour of one such fragment is a consequence of stable curvature of the helix axis in solution. The molecules involved tend to contain oligo(dA)-oligo(dT) runs which are approximately in-phase with the helix repeat; however, the precise structural elements responsible for DNA curvature have not been identified. One popular model for curvature invokes a non-coplanar 'wedge-like' conformation of ApA/TpT dinucleotide pairs. Despite a lack of direct evidence in support of this model, it has been used to provide quantitative estimates of curvature. To critically evaluate the ApA wedge model, we have performed an electrophoretic analysis of a series of closely related DNA polymers in which oligo(dA)-oligo(dT) runs of different polarity were compared. We conclude that ApA dinucleotide wedges cannot account for DNA curvature. Therefore, quantitative estimates for ApA wedge deformations, based solely on apparent curvature, cannot be correct.
来自原核生物和真核生物的DNA都产生了限制片段,这些片段在聚丙烯酰胺凝胶上进行电泳时表现出明显异常的电泳行为(迁移率降低)。我们之前已经表明,其中一个这样的片段的异常电泳行为是溶液中螺旋轴稳定弯曲的结果。所涉及的分子往往含有与螺旋重复序列大致同相的寡聚(dA)-寡聚(dT)序列;然而,尚未确定负责DNA弯曲的精确结构元件。一种流行的弯曲模型涉及ApA/TpT二核苷酸对的非共面“楔形”构象。尽管缺乏支持该模型的直接证据,但它已被用于提供弯曲度的定量估计。为了严格评估ApA楔形模型,我们对一系列密切相关的DNA聚合物进行了电泳分析,比较了不同极性的寡聚(dA)-寡聚(dT)序列。我们得出结论,ApA二核苷酸楔形不能解释DNA弯曲。因此,仅基于表观弯曲度对ApA楔形变形的定量估计不可能是正确的。