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首页> 外文期刊>Molecular and Cellular Biology >Cloning and sequence analysis of the Saccharomyces cerevisiae RAD9 gene and further evidence that its product is required for cell cycle arrest induced by DNA damage.
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Cloning and sequence analysis of the Saccharomyces cerevisiae RAD9 gene and further evidence that its product is required for cell cycle arrest induced by DNA damage.

机译：啤酒酵母RAD9基因的克隆和序列分析，并进一步证明其产物是DNA损伤诱导的细胞周期停滞所必需的。

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摘要

Procaryotic and eucaryotic cells possess mechanisms for arresting cell division in response to DNA damage. Eucaryotic cells arrest division in the G2 stage of the cell cycle, and various observations suggest that this arrest is necessary to ensure the completion of repair of damaged DNA before the entry of cells into mitosis. Here, we provide evidence that the Saccharomyces cerevisiae RAD9 gene, mutations of which confer sensitivity to DNA-damaging agents, is necessary for the cell cycle arrest phenomenon. Our studies with the rad9 delta mutation show that RAD9 plays a role in the cell cycle arrest of methyl methanesulfonate-treated cells and is absolutely required for the cell cycle arrest in the temperature-sensitive cdc9 mutant, which is defective in DNA ligase. At the restrictive temperature, cell cycle progression of cdc9 cells is blocked sometime after the DNA chain elongation step, whereas cdc9 rad9 delta cells do not arrest at this point and undergo one or two additional divisions. Upon transfer from the restrictive to the permissive temperature, a larger proportion of the cdc9 cells than of the cdc9 rad9 delta cells forms viable colonies, indicating that RAD9-mediated cell cycle arrest allows for proper ligation of DNA breaks before the entry of cells into mitosis. The rad9 delta mutation does not affect the frequency of spontaneous or UV-induced mutation and recombination, suggesting that RAD9 is not directly involved in mutagenic or recombinational repair processes. The RAD9 gene encodes a transcript of approximately 4.2 kilobases and a protein of 1,309 amino acids of Mr 148,412. We suggest that RAD9 may be involved in regulating the expression of genes required for the transition from G2 to mitosis.
机译：原核和真核细胞具有响应DNA损伤而阻止细胞分裂的机制。真核细胞在细胞周期的G2期阻滞分裂，各种观察结果表明，这种阻滞是确保细胞进入有丝分裂之前确保受损DNA修复完成所必需的。在这里，我们提供的证据表明，酿酒酵母RAD9基因的突变赋予了对DNA破坏剂的敏感性，是细胞周期停滞现象所必需的。我们对rad9 delta突变的研究表明，RAD9在甲磺酸甲酯处理的细胞的细胞周期停滞中发挥作用，并且是温度敏感的cdc9突变体中细胞周期停滞所绝对需要的，而该突变体在DNA连接酶中是有缺陷的。在限制性温度下，cdc9细胞的细胞周期进程在DNA链延长步骤后的某个时间被阻止，而cdc9 rad9 delta细胞此时不停滞并经历一到两个额外的分裂。从限制温度转移到允许温度时，cdc9细胞中比cdc9 rad9 delta细胞更大的比例形成了活菌落，这表明RAD9介导的细胞周期阻滞允许在细胞进入有丝分裂之前进行适当的DNA断裂连接。 。 rad9 delta突变不会影响自发或紫外线诱导的突变和重组的频率，这表明RAD9并不直接参与诱变或重组修复过程。 RAD9基因编码约4.2千个碱基的转录本和148,412先生的1,309个氨基酸的蛋白质。我们建议RAD9可能参与调节从G2过渡到有丝分裂所需的基因表达。

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