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在牛淋巴细胞有丝分裂刺激过程中观察到的核苷酸和碱基DNA切除修复的差异。

Differences in nucleotide and base DNA excision repair observed during mitogenic stimulation of bovine lymphocytes.

作者信息

Schrader T J

机构信息

Department of Biochemistry, University of Western Ontario, London, Canada.

出版信息

Mutat Res. 1992 Jan;273(1):29-42. doi: 10.1016/0921-8777(92)90047-7.

Abstract

The bromodeoxyuridine density-shift technique was used to examine nucleotide and base DNA excision repair in quiescent and lectin stimulated bovine lymphocytes damaged with either ultraviolet light or dimethyl sulfate (DMS). Compared to a number of human cell lines, quiescent lymphocytes were less proficient in the repair of both types of damage. Repair replication was enhanced upon mitogenic stimulation, but both the amount and time course of the increase in repair depended upon the damaging agent used. A 2-3-fold increase in UV light induced repair replication occurred early during stimulation and subsided only gradually as stimulation proceeded. However, the profile of DMS induced repair increased 7-fold and then decreased, in parallel with measurements of lectin-stimulated DNA replication. Estimates of average repair patch sizes showed that quiescent lymphocytes produced smaller patches of 7 nucleotides in response to DMS damage while UV light irradiation resulted in repair patches of 20 nucleotides. During stimulation, patch sizes appeared to increase to maximum values of 45 and 33 nucleotides in response to UV light and DMS, respectively, one day prior to the peak of DNA replication. These increases in patch size were followed by a gradual decrease towards unstimulated levels. However, the appearance of a DNA species of intermediate density in the gradient profiles made the interpretation of repair patch sizes in stimulated cells difficult. These results are discussed as evidence not only for differences in the mechanisms of nucleotide and base excision repair but also for changes in repair as the cell progresses through the cell cycle.

摘要

采用溴脱氧尿苷密度转移技术,检测静止和凝集素刺激的牛淋巴细胞在受到紫外线或硫酸二甲酯(DMS)损伤后的核苷酸和碱基DNA切除修复情况。与多种人类细胞系相比,静止淋巴细胞对这两种损伤的修复能力较差。有丝分裂原刺激可增强修复复制,但修复增加的量和时间进程取决于所使用的损伤剂。紫外线诱导的修复复制在刺激早期增加2 - 3倍,且仅在刺激持续进行时逐渐消退。然而,DMS诱导的修复情况是先增加7倍然后下降,与凝集素刺激的DNA复制测量结果平行。对平均修复片段大小的估计表明,静止淋巴细胞对DMS损伤产生的较小片段为7个核苷酸,而紫外线照射导致的修复片段为20个核苷酸。在刺激过程中,片段大小在DNA复制高峰前一天分别增加到紫外线和DMS刺激下的最大值45和33个核苷酸。这些片段大小的增加之后逐渐降至未刺激水平。然而,梯度图谱中中等密度DNA种类的出现使得对刺激细胞中修复片段大小的解释变得困难。这些结果不仅作为核苷酸和碱基切除修复机制差异的证据进行了讨论,也作为细胞在细胞周期进程中修复变化的证据进行了讨论。

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