Wellinger R J, Wolf A J, Zakian V A
Fred Hutchinson Cancer Research Center, Seattle, Washington.
Cell. 1993 Jan 15;72(1):51-60. doi: 10.1016/0092-8674(93)90049-v.
Saccharomyces telomeres consist of approximately 300 bp of C1-3A/TG1-3 DNA. Nondenaturing Southern hybridization, capable of detecting approximately 60 to approximately 300 bases of TG1-3 DNA, revealed that yeast telomeres acquired and lost TG1-3 tails, the predicted intermediate in telomere replication, in a cell cycle-dependent manner. TG1-3 tails were also detected on the ends of a linear plasmid isolated from late S phase cells. In addition, a nonlinear form of this plasmid was detected: this structure migrated in two-dimensional agarose gels like a nicked circle of the same size as the linear plasmid, but had considerably more single-stranded character than a conventional nicked circle. The evidence indicates that these circles were formed by telomere-telomere interactions involving the TG1-3 tails. These data provide evidence for a cell cycle-dependent change in telomere structure and demonstrate that TG1-3 tails, generated during replication of a linear plasmid in vivo, are capable of mediating telomere-telomere interactions.
酿酒酵母的端粒由大约300 bp的C1-3A/TG1-3 DNA组成。非变性Southern杂交能够检测大约60至大约300个碱基的TG1-3 DNA,结果显示酵母端粒以细胞周期依赖性方式获得和丢失TG1-3尾,这是端粒复制过程中预测的中间体。在从S期晚期细胞中分离出的线性质粒的末端也检测到了TG1-3尾。此外,还检测到了该质粒的一种非线性形式:这种结构在二维琼脂糖凝胶中的迁移方式类似于与线性质粒大小相同的切口环,但比传统的切口环具有更多的单链特征。有证据表明这些环是由涉及TG1-3尾的端粒-端粒相互作用形成的。这些数据为端粒结构的细胞周期依赖性变化提供了证据,并证明在体内线性质粒复制过程中产生的TG1-3尾能够介导端粒-端粒相互作用。