Kutsyĭ M P, Zakrzhevskaia D T, Gaziev A I
Biokhimiia. 1984 May;49(5):842-9.
A DNA-protein complex resistant to 8 M urea and 0.1% SDS was obtained by chromatography of nuclear matrix lysate from Ehrlich ascite carcinoma cells on Sepharose 2BCL. Separation of the complex under more severe conditions (4 M guanidine hydrochloride, 5 M urea) on hydroxylapatite resulted in protein and DNA fractions, as well as in two fractions of the DNA-protein complex. One of the fractions of this complex was enriched with single-stranded DNA and contained a 5-fold excess of newly synthesized DNA over the DNA present in the original complex. The fractions isolated from the nuclear matrix of control Ehrlich ascite carcinoma cells and the cells incubated with novobiocin revealed quantitative and qualitative differences in the electrophoretic patterns of the proteins. Treatment of cells with novobiocin resulted in inhibition of DNA replicative synthesis and an increase in the protein/DNA ratio in the nuclear matrix.
通过将艾氏腹水癌细胞的核基质裂解物在琼脂糖2BCL上进行层析,获得了一种对8M尿素和0.1%十二烷基硫酸钠有抗性的DNA-蛋白质复合物。在更严苛的条件下(4M盐酸胍、5M尿素),该复合物在羟基磷灰石上分离得到蛋白质和DNA组分,以及两个DNA-蛋白质复合物组分。该复合物的其中一个组分富含单链DNA,且新合成的DNA比原始复合物中的DNA多出5倍。从对照艾氏腹水癌细胞的核基质以及与新生霉素一起孵育的细胞中分离得到的组分,在蛋白质的电泳图谱上显示出定量和定性的差异。用新生霉素处理细胞会导致DNA复制性合成受到抑制,并且核基质中蛋白质/DNA的比例增加。