Radic M Z, Lundgren K, Hamkalo B A
Cell. 1987 Sep 25;50(7):1101-8. doi: 10.1016/0092-8674(87)90176-0.
Cloned, sequenced mouse satellite DNA exhibits properties characteristic of molecules that possess a stable curvature. Circularly permuted fragments containing the region predicted to bend were used to map the curvature relative to DNA sequence. The altered mobility of these fragments in polyacrylamide gels is reversed when gels are run in the presence of distamycin A, a drug that binds preferentially to AT-rich DNA. Treatment of living mouse cells with this drug dramatically reduces the condensation of centromeric heterochromatin, the exclusive location of satellite sequences. In situ hybridization of satellite probes to extended chromosomes at the electron microscope level shows that satellite does not comprise a single block but is distributed throughout the centromere region. Based on these experiments, we hypothesize that the structure of mouse satellite DNA is an important feature of centromeric heterochromatin condensation.
克隆并测序的小鼠卫星DNA展现出具有稳定曲率的分子的特征特性。含有预计会弯曲区域的环状排列片段被用于绘制相对于DNA序列的曲率图谱。当在偏端霉素A存在的情况下进行凝胶电泳时,这些片段在聚丙烯酰胺凝胶中改变的迁移率会逆转,偏端霉素A是一种优先结合富含AT的DNA的药物。用这种药物处理活的小鼠细胞会显著减少着丝粒异染色质的凝聚,而卫星序列仅位于该区域。在电子显微镜水平下,将卫星探针与伸展的染色体进行原位杂交表明,卫星DNA并非构成一个单一的区域,而是分布在整个着丝粒区域。基于这些实验,我们推测小鼠卫星DNA的结构是着丝粒异染色质凝聚的一个重要特征。