S'iakste N I, Zaboĭkin M M, Erenpreĭsa E A, Likhtenshteĭn A V, Shapot V S
Mol Biol (Mosk). 1985 Sep-Oct;19(5):1231-41.
There are two types of DNA-nuclear matrix interactions in animal cells as revealed by the release of DNA from isolated nuclei by three successive gradients: NaCl, LiCl-urea and temperature. Nuclei were treated with dissociating agents while being adsorbed on the Celite columns. "Weak" DNA-matrix interactions which dissociate in 1.5 M LiCl-3 M urea at 2 degrees appear to be sensitive to ethidium bromide and resistant to exogeneous nucleases (DNAase I, DNAase II and micrococcal nuclease), to DNA-damaging agents, including alkylators and gamma-irradiation, and also to psoralen-induced cross-links. "Strong" DNA-matrix interactions proved to be very different. They dissociate in 4 M LiCl-8 M urea at approximately 90 degrees, are very sensitive to DNAase I and other nucleases, slightly sensitive to chemicals and irradiation at doses stimulating single-stranded DNA breaks, but resistant to ethidium bromide. DNA strand separation seems to be necessary prerequisite for DNA release from its "strong" complex with nuclear matrix. A model for the topological link between DNA and the nuclear matrix involved in DNA replication complex is discussed.
通过用氯化钠、氯化锂-尿素和温度这三种连续梯度从分离的细胞核中释放DNA,揭示了动物细胞中存在两种类型的DNA-核基质相互作用。细胞核在吸附于硅藻土柱上时用解离剂处理。在2℃下于1.5M氯化锂-3M尿素中解离的“弱”DNA-基质相互作用似乎对溴化乙锭敏感,对外源核酸酶(DNA酶I、DNA酶II和微球菌核酸酶)、对包括烷化剂和γ射线在内的DNA损伤剂以及对补骨脂素诱导的交联具有抗性。“强”DNA-基质相互作用则非常不同。它们在约90℃下于4M氯化锂-8M尿素中解离,对DNA酶I和其他核酸酶非常敏感,对刺激单链DNA断裂的剂量的化学物质和辐射略有敏感,但对溴化乙锭具有抗性。DNA链分离似乎是DNA从其与核基质的“强”复合物中释放的必要前提。讨论了参与DNA复制复合物的DNA与核基质之间拓扑连接的模型。