Møllegaard Niels Erik, Nielsen Peter E
Department of Medical Biochemistry and Genetics, The Panum Institute, University of Copenhagen, Blegdamsvej 3c, 2200 Copenhagen N, Denmark.
Biochemistry. 2003 Jul 22;42(28):8587-93. doi: 10.1021/bi0343267.
DNA curvature is affected by elevated temperature and dehydrating agents such as 2-methyl-2,4-pentanediol (MPD) (used in crystallization). This effect of MPD has been ascribed to a specific distortion of the structure of adenine tracts (A-tracts), probably through a deformation of the characteristic narrow minor groove. Uranyl photoprobing indicates that a narrowed minor groove is present in all A/T regions containing four or more A/T base pairs. Consequently, this technique may be employed to study conformational changes in other A/T-rich sequences than pure A-tracts. In this study we use uranyl photoprobing to demonstrate that the effect of elevated temperature and MPD is analogous on both "normal" and curve-inducing A/T-rich sequences. The results therefore indicate that under these conditions the minor groove is widened in all A/T sequences and not only in pure A-tracts as previously suggested. Thus, the rather subtle structural difference of AT regions and A-tracts in nonbent DNA versus A-tracts in bent DNA may be quantitative rather than qualitative; i.e., the structure is more persistent and/or rigid in bent DNA.
DNA 弯曲度受高温以及脱水剂(如用于结晶的 2-甲基-2,4-戊二醇(MPD))影响。MPD 的这种作用被归因于腺嘌呤序列(A 序列)结构的特定扭曲,可能是通过特征性窄小沟的变形。铀酰光探测表明,在所有包含四个或更多 A/T 碱基对的 A/T 区域都存在窄小沟。因此,该技术可用于研究除纯 A 序列之外的其他富含 A/T 序列的构象变化。在本研究中,我们使用铀酰光探测来证明高温和 MPD 对“正常”和诱导弯曲的富含 A/T 序列的影响是相似的。因此,结果表明在这些条件下,所有 A/T 序列中的小沟都会变宽,而不仅仅是如先前所认为的仅在纯 A 序列中。因此,在未弯曲 DNA 中 A/T 区域和 A 序列与弯曲 DNA 中 A 序列之间相当细微的结构差异可能是数量上的而非质量上的;即,弯曲 DNA 中的结构更持久和/或更刚性。