Kypr J, Penázová H, Sági J, Pospísilová S, Vorlícková M
Institute of Biophysics, Czech Academy of Sciences, Brno.
J Biomol Struct Dyn. 1994 Jun;11(6):1225-36. doi: 10.1080/07391102.1994.10508065.
Alkaline agarose gel electrophoresis was used to detect UV-induced crosslinking of the strands of poly(dA-dT) and related alternating purine-pyrimidine DNAs in solutions stabilizing various polynucleotide conformations. Strands of the B-form and A-form of poly(dA-dT) were not crosslinked but a UV dose-dependent retarded species appeared in the denaturing gels in parallel with the polynucleotide isomerization into the unusual X-form. Most other polynucleotides adopting the X-form were crosslinked as well. The exceptions include the X-forms of poly(dA-butyl5dU) and poly(dA-pentyl5dU) whose strands do not crosslink because the long exocyclic substituents attached to uracil make the photodimerization impossible. Strands of poly(amino2dA-dT) and poly(dA, amino2dA-dT), the latter polynucleotide containing roughly equal amounts of amino2adenine and adenine, also do not crosslink upon UV irradiation because they isomerize into an A-like conformation which is different from the X-form of poly (dA-dT). In contrast, strands of the mixed copolymers of poly(dA, amino2dA-dT) containing low amino2adenine contents are crosslinked upon UV irradiation, in accordance with the observation that they isomerize into the X-form.
碱性琼脂糖凝胶电泳用于检测在稳定各种多核苷酸构象的溶液中紫外线诱导的聚(dA-dT)链以及相关交替嘌呤 - 嘧啶DNA链的交联。聚(dA-dT)的B型和A型链未发生交联,但在变性凝胶中出现了一种紫外线剂量依赖性的滞后物种,同时多核苷酸异构化为不寻常的X型。大多数采用X型的其他多核苷酸也发生了交联。例外情况包括聚(dA-丁基5dU)和聚(dA-戊基5dU)的X型,其链不发生交联,因为连接到尿嘧啶上的长环外取代基使光二聚化无法进行。聚(氨基2dA-dT)和聚(dA,氨基2dA-dT)的链,后者多核苷酸含有大致等量的氨基2腺嘌呤和腺嘌呤,在紫外线照射下也不发生交联,因为它们异构化为一种类似A的构象,这与聚(dA-dT)的X型不同。相比之下,含有低氨基2腺嘌呤含量的聚(dA,氨基2dA-dT)混合共聚物的链在紫外线照射下发生交联,这与它们异构化为X型的观察结果一致。