研究芽殖酵母中复制叉崩溃的方法。
Methods to study replication fork collapse in budding yeast.
作者信息
Liberi Giordano, Cotta-Ramusino Cecilia, Lopes Massimo, Sogo Jose', Conti Chiara, Bensimon Aaron, Foiani Marco
机构信息
F.I.R.C. Institute of Molecular Oncology Foundation and DSBB-University of Milan, Milan, Italy.
出版信息
Methods Enzymol. 2006;409:442-62. doi: 10.1016/S0076-6879(05)09026-9.
Replication of the eukaryotic genome is a difficult task, as cells must coordinate chromosome replication with chromatin remodeling, DNA recombination, DNA repair, transcription, cell cycle progression, and sister chromatid cohesion. Yet, DNA replication is a potentially genotoxic process, particularly when replication forks encounter a bulge in the template: forks under these conditions may stall and restart or even break down leading to fork collapse. It is now clear that fork collapse stimulates chromosomal rearrangements and therefore represents a potential source of DNA damage. Hence, the comprehension of the mechanisms that preserve replication fork integrity or that promote fork collapse are extremely relevant for the understanding of the cellular processes controlling genome stability. Here we describe some experimental approaches that can be used to physically visualize the quality of replication forks in the yeast S. cerevisiae and to distinguish between stalled and collapsed forks.
真核生物基因组的复制是一项艰巨的任务,因为细胞必须协调染色体复制与染色质重塑、DNA重组、DNA修复、转录、细胞周期进程以及姐妹染色单体黏连。然而,DNA复制是一个潜在的基因毒性过程,尤其是当复制叉遇到模板中的凸起时:在这些条件下,复制叉可能会停滞并重新启动,甚至崩溃导致复制叉瓦解。现在很清楚,复制叉瓦解会刺激染色体重排,因此代表了DNA损伤的一个潜在来源。因此,理解维持复制叉完整性或促进复制叉瓦解的机制对于理解控制基因组稳定性的细胞过程极为重要。在这里,我们描述了一些实验方法,可用于直观地观察酿酒酵母中复制叉的质量,并区分停滞的和瓦解的复制叉。