在酵母的双链断裂修复过程中,Srs2和Sgs1-Top3抑制交叉
Grzegorz Ira1, Anna Malkova, Giordano Liberi
1Rosenstiel Center and Department of Biology, Brandeis University, Waltham, MA 02454, USA.
Cell
|November 19, 2003
概括
酵母中的遗传重组是受调节的,有不同的修复途径影响交叉结果. 像SGS1和SRS2这样的关键蛋白调节交叉频率,影响遗传稳定性.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 细胞生物学 细胞生物学
背景情况:
- 线粒细胞中的同源重组对于遗传稳定至关重要.
- 基因转换和交叉事件之间的关系尚未完全理解.
- 监管交叉结果对于防止基因组不稳定性至关重要.
研究的目的:
- 调查平芽酵母菌中同源重组和交叉的调节.
- 确定在DNA修复过程中影响交叉结果的特定因素.
- 阐明区分交叉和非交叉维修路径的机制.
主要方法:
- 基因转换和交叉事件在平芽酵母中的分析.
- 关键蛋白质的基因操纵,包括SGS1 (BLM/WRN同类) 和SRS2酶.
- 研究基因删除和过度表达对重组途径的影响.
- 研究基因转换事件与交叉事件相关的时间.
主要成果:
- 没有交叉的基因转换比交叉的基因转换早30分钟,这表明了不同的修复机制.
- 删除SGS1或SRS2会使交叉频率增加2到3倍.
- 过度表达SRS2显著减少交叉事件.
- 在srs2Δ细胞中RAD51的过度表达抑制了非交叉重组途径.
结论:
- Sgs1和Top3可能通过解决双休日结的中间体来减少交叉.
- Srs2促进了合成依赖的链化 (SDSA) 途径,有利于非交叉结果.
- 这些发现突出了复杂的调节的重组途径,以保持遗传稳定.
相关概念视频
DNA Topoisomerases
Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types. Type I...
Types and Mechanism of action
Topoisomerases are divided into two main types. Type I...
Fixing Double-strand Breaks
The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
Homologous Recombination
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
Restarting Stalled Replication Forks
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
Fixing Double-strand Breaks
The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...


