Collins I, Newlon C S
Department of Microbiology and Molecular Genetics, UMD-New Jersey Medical School, Newark 07103.
Mol Cell Biol. 1994 May;14(5):3524-34. doi: 10.1128/mcb.14.5.3524-3534.1994.
Autonomously replicating sequence (ARS) elements are identified by their ability to promote high-frequency transformation and extrachromosomal replication of plasmids in the yeast Saccharomyces cerevisiae. Six of the 14 ARS elements present in a 200-kb region of Saccharomyces cerevisiae chromosome III are mitotic chromosomal replication origins. The unexpected observation that eight ARS elements do not function at detectable levels as chromosomal replication origins during mitotic growth suggested that these ARS elements may function as chromosomal origins during premeiotic S phase. Two-dimensional agarose gel electrophoresis was used to map premeiotic replication origins in a 100-kb segment of chromosome III between HML and CEN3. The pattern of origin usage in premeiotic S phase was identical to that in mitotic S phase, with the possible exception of ARS308, which is an inefficient mitotic origin associated with CEN3. CEN3 was found to replicate during premeiotic S phase, demonstrating that the failure of sister chromatids to disjoin during the meiosis I division is not due to unreplicated centromeres. No origins were found in the DNA fragments without ARS function. Thus, in both mitosis and meiosis, chromosomal replication origins are coincident with ARS elements but not all ARS elements have chromosomal origin function. The efficiency of origin use and the patterns of replication termination are similar in meiosis and in mitosis. DNA replication termination occurs over a broad distance between active origins.
自主复制序列(ARS)元件是通过其促进质粒在酿酒酵母中高频转化和染色体外复制的能力来鉴定的。酿酒酵母第三条染色体200 kb区域中的14个ARS元件中有6个是有丝分裂染色体复制起点。一个意外的观察结果是,在有丝分裂生长期间,8个ARS元件作为染色体复制起点在可检测水平上不起作用,这表明这些ARS元件可能在减数分裂前S期作为染色体起点发挥作用。二维琼脂糖凝胶电泳用于绘制HML和CEN3之间第三条染色体100 kb片段中的减数分裂前复制起点。减数分裂前S期的起点使用模式与有丝分裂S期相同,可能ARS308除外,它是一个与CEN3相关的低效有丝分裂起点。发现CEN3在减数分裂前S期进行复制,这表明在减数第一次分裂期间姐妹染色单体不能分离不是由于着丝粒未复制。在没有ARS功能的DNA片段中未发现起点。因此,在有丝分裂和减数分裂中,染色体复制起点都与ARS元件重合,但并非所有ARS元件都具有染色体起点功能。减数分裂和有丝分裂中起点使用效率和复制终止模式相似。DNA复制终止发生在活跃起点之间的广泛距离上。