Kobayashi T, Horiuchi T
Laboratory of Gene Expression and Regulation, National Institute for Basic Biology, Okazaki, Japan.
Genes Cells. 1996 May;1(5):465-74. doi: 10.1046/j.1365-2443.1996.d01-256.x.
In the rRNA gene cluster on the Saccharomyces cerevisiae chromosome XII, a unique site that blocks progression of the replication fork exists in a single unit of the repeats. The site RFB (replication fork blocking) is located within one of two cis-elements present in a nontranscribed region of each repeated unit, which are required for obtaining maximal activity of a hotspot (HOT1) dependent recombination.
To investigate the correlation between replication fork blocking at RFB and homologous recombination at HOT1, we have isolated Hot1-defective mutants and examined their ability for fork blocking at RFB. Amongst 23 isolated mutants, four were found to be defective in both abilities. Genetic analysis of the mutants reveals that a single mutation, named fob1 (fork blocking less), is responsible for the defects in both abilities. The FOB1 gene is located on chromosome IV and has no homology with any other genes listed in DNA data banks.
The pleiotropic effect of the fob1 mutation suggests that homologous recombination at HOT1 is closely linked with DNA replication fork blocking event at RFB.
在酿酒酵母十二号染色体上的核糖体RNA基因簇中,重复序列的单个单元内存在一个阻止复制叉前进的独特位点。RFB(复制叉阻断)位点位于每个重复单元非转录区域中存在的两个顺式元件之一内,这两个顺式元件是获得依赖热点(HOT1)的重组最大活性所必需的。
为了研究RFB处的复制叉阻断与HOT1处的同源重组之间的相关性,我们分离了Hot1缺陷型突变体,并检测了它们在RFB处的叉阻断能力。在分离出的23个突变体中,发现有4个在这两种能力上都存在缺陷。对这些突变体的遗传分析表明,一个名为fob1(叉阻断减少)的单一突变导致了这两种能力的缺陷。FOB1基因位于四号染色体上,与DNA数据库中列出的任何其他基因均无同源性。
fob1突变的多效性效应表明,HOT1处的同源重组与RFB处的DNA复制叉阻断事件密切相关。