哺乳动物间期细胞核中复制DNA与核基质附着的分析。
Analysis of the attachment of replicating DNA to a nuclear matrix in mammalian interphase nuclei.
作者信息
Dijkwel P A, Mullenders L H, Wanka F
出版信息
Nucleic Acids Res. 1979 Jan;6(1):219-30. doi: 10.1093/nar/6.1.219.
Abstract
The attachment of replicating DNA to a rapidly sedimenting nuclear structure was investigated by digestion with various nucleases. When DNA was gradually removed by DNase I, pulse label incorporated during either 1 min or during 1 hour in the presence of arabinosylcytosine, remained preferentially attached to the nuclear structure. Single strand specific digestion by nuclease S1 or staphylococcal nuclease at low concentrations caused a release of about 30% of the pulse label, without significantly affecting the attachment of randomly labelled DNA. The released material had a low sedimentation coefficient and contained most of the Okasaki fragments. The remaining pulse label was less accessible to further digestion by double strand specific nuclease activity than the bulk DNA. The results suggest that an attachment of the replication fork to the nuclear structure occurs at sites behind but close to the branch point.
摘要
通过用各种核酸酶消化来研究复制中的DNA与快速沉降的核结构的附着情况。当用DNase I逐渐去除DNA时，在阿拉伯糖胞苷存在下1分钟或1小时内掺入的脉冲标记物仍优先附着于核结构。低浓度的核酸酶S1或葡萄球菌核酸酶进行单链特异性消化导致约30%的脉冲标记物释放，而对随机标记的DNA的附着没有显著影响。释放的物质沉降系数低，并且包含大多数冈崎片段。剩余的脉冲标记物比大部分DNA更不易被双链特异性核酸酶活性进一步消化。结果表明复制叉与核结构的附着发生在分支点后方但靠近分支点的位点。
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