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Correlation of sister chromatid exchange formation through homologous recombination with ribonucleotide reductase inhibition.

We conducted the recombination and sister chromatid exchange (SCE) assays with five chemicals (hydroxyurea (HU), resveratrol, 4-hydroxy-trans-stilbene, 3-hydroxy-trans-stilbene, and mitomycin C) in Chinese hamster cell line SPD8/V79 to confirm directly that SCE is a result of homologous recombination (HR). SPD8 has a partial duplication in exon 7 of the endogenous hprt gene and can revert to wild type by homologous recombination. All chemicals were positive in both assays except for 3-hydroxy-trans-stilbene, which was negative in both. HU, resveratrol, and 4-hydroxy-trans-stilbene were scavengers of the tyrosyl free radical of the R2 subunit of mammalian ribonucleotide reductase. Tyrosyl free radical scavengers disturb normal DNA replication, causing replication fork arrest. Mitomycin C is a DNA cross-linking agent that also causes replication fork arrest. The present study suggests that replication fork arrest, which is similar to the early phases of HR, leads to a high frequency of recombination, resulting in SCEs. The findings show that SCE may be mediated by HR.

In this paper we show that sister chromatid exchanges (SCE) can be analysed by computer-supported determination of areas and shapes of darkly stained chromatid sections. In combination with appropriate statistical analyses, our method leads to measurements of SCE frequencies in neocarzinostatin-treated Chinese hamster ovary cells which are nearly as precise as the ones obtained by classical microscopical analyses. We discuss our data as an approach to the development of a fully automated SCE scoring system.

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  • 1 0 年前
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    相同染色體交換形成經由同源重組與核糖核甘酸還原脢抑制物相互作用。

    我們在中國蒼鼠細胞株SPD8/V79作同源重組和染色體交換分析五種化學物(烴基尿hydroxyurea,白藜蘆醇resveratrol,4氫氧根對二苯代乙烯4-hydroxy-trans-stilbene,3氫氧根對二苯代乙烯3-hydroxy-trans-stilbene和 絲裂黴素C mitomycin C),可以立即確定染色體交換是同源重組的結果。在內部基因突變的外顯子7 (exon 7)中,SPD8可以部分複製,並且可以經由同源重組把蒼鼠回復為野生種類。在同源重組和染色體交換分析中,除了3氫氧根對二苯代乙烯呈現陰性反應外,所有化學物皆呈現陽性反應。烴基尿hydroxyurea,白藜蘆醇resveratrol,4氫氧根對二苯代乙烯4-hydroxy-trans-stilbene是哺乳動物核糖核甘酸還原脢R2次單體酪氨酰根游離基(tyrosy1 free radical)的清除劑。根游離基清除劑會干擾正常的DNA複製,造成複製交叉停止。絲裂黴素C是一種DNA交叉結合劑,會引起複製交叉停止。現今研究提出複製交叉停止與初期的同源重組類似,致使高頻率的同源重組,導致染色體交換。這個研究顯示同源重組可能促成染色體交換。

    在此報告中,我們說明分析相同染色體交換能經由電腦證實區位決定和深色染色體區域的形狀。與適當的數據分析結合,我們的方法能在經處理的新抑癌蛋白的中國蒼鼠卵巣細胞中,測量相同染色體交換次數,這個測量的準確度近於在傳統顯微鏡的分析。我們將討論我們的數據作為一種完全自動相同染色體交換記號系統發展的方法。

    參考資料: me
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