de Massy B, Nicolas A
Institut de Génétique et Microbiologie, Université Paris-Sud, Orsay, France.
EMBO J. 1993 Apr;12(4):1459-66. doi: 10.1002/j.1460-2075.1993.tb05789.x.
During meiosis, a transient DNA double-strand break (DSB) occurs in the promoter region (positions -200/-185) of the Saccharomyces cerevisiae ARG4 gene and is a likely intermediate in the initiation of meiotic gene conversion events in this region. We report here a functional analysis of the ARG4 DSB based on the study of various deletions in this chromosomal region. We have identified several cis-acting elements located within the -465/+3 region of the ARG4 promoter that control the formation of this DSB. The -465/-317 region includes a transcription terminator and is necessary for a normal amount of ARG4 DSB, but not for its positioning. The -316/-140 region can be replaced by an unrelated DNA sequence where a meiotic DSB then occurs, suggesting that the site of DSB is not sequence-specific, but is positioned at a fixed distance from the adjacent -139/+3 region. Also, in all strains constructed, the amount of meiotic DSB is correlated with the frequency of gene conversion in ARG4, which provides a strong argument for the initiation of gene conversion by a DSB in this region of the yeast genome.
在减数分裂过程中,酿酒酵母ARG4基因的启动子区域(位置-200/-185)会发生短暂的DNA双链断裂(DSB),这可能是该区域减数分裂基因转换事件起始过程中的一个中间体。我们在此基于对该染色体区域各种缺失的研究报告了ARG4 DSB的功能分析。我们已经鉴定出位于ARG4启动子-465/+3区域内的几个顺式作用元件,它们控制着这种DSB的形成。-465/-317区域包含一个转录终止子,对于正常数量的ARG4 DSB是必需的,但对于其定位不是必需的。-316/-140区域可以被一个不相关的DNA序列取代,在此处会发生减数分裂DSB,这表明DSB的位点不是序列特异性的,而是位于距相邻的-139/+3区域固定距离处。此外,在构建的所有菌株中,减数分裂DSB的数量与ARG4中基因转换的频率相关,这为酵母基因组该区域中由DSB引发基因转换提供了有力证据。