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使用不同γ射线剂量率诱导酿酒酵母中DNA双链断裂的修复:脉冲场凝胶电泳分析

Repair of DNA double-strand breaks induced in Saccharomyces cerevisiae using different gamma-ray dose-rates: a pulsed-field gel electrophoresis analysis.

作者信息

Dardalhon M, Nohturfft A, Meniel V, Averbeck D

机构信息

Institut Curie-Biologie, CNRS URA 1292, Paris, France.

出版信息

Int J Radiat Biol. 1994 Mar;65(3):307-14. doi: 10.1080/09553009414550361.

Abstract

We investigated the effects of gamma-ray exposures at high dose-rate (HDR, 23.2 Gy/min) and low dose-rate (LDR, 0.47 Gy/min) on survival and the induction of DNA double-strand breaks (dsb) in a diploid wild-type (D7) and the repair-deficient mutant strain rad52/rad52 of Saccharomyces cerevisiae. Analysis by pulsed-field gel electrophoresis (PFGE) using a contour homogeneous electric field apparatus revealed that, at HDR, in the range 0-400 Gy, dsb are induced as a linear function of gamma-ray dose. Liquid holding recovery in non-nutrient medium (LHR) for 48 h of wild-type cells treated at HDR, significantly increased survival and reduced the yield of dsb. Such changes did not occur in rad52/rad52 cells defective in the repair of dsb. Thus, in gamma-irradiated wild-type cells, an efficient repair of dsb is taking place during LHR. Treatments of wild-type cells at LDR resulted in higher survival and an approximately two-fold lower yield of dsb than at HDR. Such a dose-rate effect was absent in rad52/rad52 cells suggesting that, in wild-type cells during LDR exposures, significant amounts of dsb can be repaired. This repair could be very much accentuated by 48-h LHR of wild-type cells treated at LDR. The relationship observed between gamma-ray survival and dsb repair clearly indicates that increases in survival of wild-type cells, during LDR as compared with HDR exposures and after LHR, are strongly related to the repair of dsb.

摘要

我们研究了高剂量率(HDR,23.2 Gy/分钟)和低剂量率(LDR,0.47 Gy/分钟)的γ射线照射对酿酒酵母二倍体野生型(D7)和修复缺陷突变株rad52/rad52的存活及DNA双链断裂(dsb)诱导的影响。使用轮廓均匀电场装置通过脉冲场凝胶电泳(PFGE)分析表明,在HDR条件下,在0 - 400 Gy范围内,dsb的诱导与γ射线剂量呈线性函数关系。在HDR处理的野生型细胞在无营养培养基中进行48小时的液体保持恢复(LHR),显著提高了存活率并降低了dsb的产量。在dsb修复存在缺陷的rad52/rad52细胞中未发生此类变化。因此,在γ射线照射的野生型细胞中,LHR期间发生了dsb的有效修复。在LDR条件下处理野生型细胞,与HDR相比,存活率更高,dsb产量降低约两倍。rad52/rad52细胞中不存在这种剂量率效应,这表明在LDR照射期间的野生型细胞中,大量dsb可以被修复。在LDR处理的野生型细胞进行48小时LHR后,这种修复作用会大大增强。γ射线存活与dsb修复之间观察到的关系清楚地表明,与HDR照射相比,在LDR期间以及LHR之后,野生型细胞存活率的提高与dsb的修复密切相关。

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