Morrow D M, Connelly C, Hieter P
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Genetics. 1997 Oct;147(2):371-82. doi: 10.1093/genetics/147.2.371.
Introduction of a chromosome fragmentation vector (CFV) into the budding yeast Saccharomyces cerevisiae results in a targeted homologous recombination event that yields an independently segregating chromosome fragment (CF) and an alteration in the strain's karyotype. Fragmentation with an acentric CFV directed in a centromere-proximal orientation generates a CF that contains all sequences proximal to the targeting segment and results in loss of the endogenous targeted chromosome to yield a 2N-1 + CF karyotype. In contrast, fragmentation with a centric CFV directed in a centromere-distal orientation generates a CF that contains all sequences distal to the targeting segment and retention of the endogenous targeted chromosome to yield a 2N + CF karyotype. We have termed this phenomenon "break copy" duplication. Using yeast strains in which the centromere had been transposed to a new location, it was demonstrated that the centromere inhibited break copy duplication. These data suggest that CF formation is the product of an unscheduled DNA replication event initiated by the free end of the CFV and is analogous to a "half" double-strand break gap-repair reaction. We suggest that break copy duplication may have evolved as a mechanism for maintenance of ploidy following DNA breakage.
将染色体片段化载体(CFV)导入出芽酵母酿酒酵母会导致靶向同源重组事件,产生一个独立分离的染色体片段(CF)并改变菌株的核型。用着丝粒近端定向的无着丝粒CFV进行片段化会产生一个CF,其包含靶向片段近端的所有序列,并导致内源性靶向染色体丢失,从而产生2N - 1 + CF核型。相比之下,用着丝粒远端定向的着丝粒CFV进行片段化会产生一个CF,其包含靶向片段远端的所有序列,并保留内源性靶向染色体,从而产生2N + CF核型。我们将这种现象称为“断裂复制”重复。使用着丝粒已转座到新位置的酵母菌株,证明着丝粒抑制断裂复制重复。这些数据表明,CF形成是由CFV的自由端引发的意外DNA复制事件的产物,类似于“半”双链断裂缺口修复反应。我们认为,断裂复制重复可能已经进化为一种在DNA断裂后维持倍性的机制。