Friedman K L, Brewer B J, Fangman W L
Genetics Department, University of Washington, Seattle 98195-7360, USA.
Genes Cells. 1997 Nov;2(11):667-78. doi: 10.1046/j.1365-2443.1997.1520350.x.
An understanding of the replication programme at the genome level will require the identification and characterization of origins of replication through large, contiguous regions of DNA. As a step toward this goal, origin efficiencies and replication times were determined for 10 ARSs spanning most of the 270 kilobase (kb) chromosome VI of Saccharomyces cerevisiae.
Chromosome VI shows a wide variation in the percentage of cell cycles in which different replication origins are utilized. Most of the origins are activated in only a fraction of cells, suggesting that the pattern of origin usage on chromosome VI varies greatly within the cell population. The replication times of fragments containing chromosome VI origins show a temporal pattern that has been recognized on other chromosomes--the telomeres replicate late in S phase, while the central region of the chromosome replicates early.
As demonstrated here for chromosome VI, analysis of the direction of replication fork movement along a chromosome and determination of replication time by measuring a period of hemimethylation may provide an efficient means of surveying origin activity over large regions of the genome.
要在基因组水平上理解复制程序,需要通过大片连续的DNA区域来鉴定和表征复制起点。作为朝着这一目标迈出的一步,我们测定了酿酒酵母270千碱基(kb)的第六条染色体上大部分区域的10个自主复制序列(ARS)的起点效率和复制时间。
第六条染色体在不同复制起点被利用的细胞周期百分比方面表现出很大差异。大多数起点仅在一部分细胞中被激活,这表明第六条染色体上起点的使用模式在细胞群体中差异很大。包含第六条染色体起点的片段的复制时间呈现出一种在其他染色体上也已被认识到的时间模式——端粒在S期后期复制,而染色体的中央区域则较早复制。
正如这里对第六条染色体所证明的那样,分析沿着染色体的复制叉移动方向以及通过测量半甲基化时期来确定复制时间,可能提供一种在基因组大片区域上检测起点活性的有效方法。