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首页> 外文期刊>Molecular and Cellular Biology >Efficient repair of HO-induced chromosomal breaks in Saccharomyces cerevisiae by recombination between flanking homologous sequences.
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Efficient repair of HO-induced chromosomal breaks in Saccharomyces cerevisiae by recombination between flanking homologous sequences.

机译：通过侧翼同源序列之间的重组，有效修复啤酒酵母中HO诱导的染色体断裂。

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摘要

Novel recombinational repair of a site-specific double-strand break (DSB) in a yeast chromosome was investigated. When the recognition site for the HO endonuclease enzyme is embedded in nonyeast sequences and placed between two regions of homology, expression of HO endonuclease stimulates recombination between the homologous flanking regions to yield a deletion, the apparent product of an intrachromosomal exchange between direct repeats. This deletion-repair event is very efficient, thus preventing essentially all the potential lethality due to the persistence of a DSB. Interestingly, unlike previous studies involving spontaneous recombination between chromosomal repeats, the recombination events stimulated by HO-induced DSBs are accompanied by loss of the sequences separating the homologous regions greater than 99.5% of the time. Repair is dependent on the RAD52 gene. The deletion-repair event provides an in vivo assay for the sensitivity of any particular recognition site to HO cleavage. By taking advantage of a galactose-inducible HO gene, it has been possible to follow the kinetics of this event at the DNA level and to search for intermediates in this reaction. Deletion-repair requires approximately 45 min and is inhibited when cycloheximide is added after HO endonuclease cleavage.
机译：研究了酵母染色体中位点特异性双链断裂（DSB）的新型重组修复。当HO核酸内切酶的识别位点嵌入非酵母序列中并置于两个同源性区域之间时，HO核酸内切酶的表达刺激同源侧翼区之间的重组产生缺失，这是直接重复之间的染色体内交换的明显产物。这种删除修复事件非常有效，因此，由于DSB的持久性，基本上可以防止所有潜在的致命性。有趣的是，与先前的涉及染色体重复之间的自发重组的研究不同，由HO诱导的DSB刺激的重组事件伴随着将同源区域分开的序列的丢失时间超过99.5％。修复取决于RAD52基因。缺失修复事件为任何特定识别位点对HO裂解的敏感性提供了一种体内测定方法。通过利用半乳糖可诱导的HO基因，可以在DNA水平上追踪该事件的动力学并在该反应中寻找中间体。缺失修复大约需要45分钟，并且在HO内切核酸酶切割后加入环己酰亚胺时会被抑制。

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