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首页> 外文学位 >ANALYSIS OF THE INDUCTION OF CHROMOSOME ABERRATIONS AT THE MOLECULAR LEVEL USING RESTRICTION ENDONUCLEASES.
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ANALYSIS OF THE INDUCTION OF CHROMOSOME ABERRATIONS AT THE MOLECULAR LEVEL USING RESTRICTION ENDONUCLEASES.

机译：使用限制性内切酶分析分子水平上的染色体畸变。

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摘要

A confounding factor in the study of radiation-induced chromosome aberration induction is the wide variety of DNA lesions that radiation produces. Restriction endonucleases (REs), which produce double strand breaks (DSBs) at specific DNA sequences, were used to study the induction of chromosome aberration induction by specific DNA lesions. Osmolytic shock of pinocytic vesicles reliably and effectively introduced REs into viable CHO cells. REs, which act as a radiomimetic agent, displayed a linear dose response for aberration induction. Cut-site frequency rather than cut-end structure appeared to determine a particular enzymes's efficiency at aberration induction.; DNA repair inhibition studies indicated RE-induced double-strand break (DSB) repair is complete within 1 hour after treatment. G{dollar}sb1{dollar} studies using cytosine arabinoside (araC) and aphidicolin indicated there is a synthesis step in the repair of RE-induced DSB. 3-aminobenzamide (3-AB), a poly(ADP-ribosyl)ation inhibitor, enhanced RE-induced aberrations during G{dollar}sb1{dollar}; its effect appears to be determined by the amount or distribution of RE-induced DSB. During G{dollar}sb2{dollar} RE treatments, araC enhanced chromatid deletion and achromatic lesion yields, but not exchange yields. 3-AB had little effect on G{dollar}sb2{dollar} induced aberrations. The DNA repair inhibition experiments suggest the presence of at least two repair mechanisms for RE-induced DSB: recombinational repair involving a synthesis step, and ligational repair regulated by poly(ADP-ribosyl)ation.; Interaction experiments involving different pairs of REs gave additional evidence for ligational repair. It appears, however, that cut-end structure affects the ability of different RE-induced DSBs to interact.; Preliminary data were obtained on the development of two different systems to study nonrandom chromosomal alterations. Studies with different mouse-hamster hybrid cell lines indicated that X-rays induced aberrations throughout the hybrid genome. Mitomycin C, however, preferentially induced aberrations in the mouse centromeric heterochromatin, which is rich in repetitive satellite DNA sequences. Not I, an enzyme with a rare recognition sequence, appeared to induce aberrations in only a few locations in the CHO karyotype. These two systems may provide a valuable tool for studying the role nonrandom chromosomal changes play in cell transformation.
机译：辐射诱导的染色体畸变诱导研究中的一个混杂因素是辐射产生的多种DNA损伤。限制性内切酶（REs）在特定的DNA序列上产生双链断裂（DSBs），用于研究特定DNA损伤对染色体畸变的诱导作用。渗透性囊泡的渗透性休克可靠而有效地将REs引入到了可行的CHO细胞中。充当放射模拟剂的稀土元素表现出线性的剂量响应，以诱导像差。切位频率而非切端结构似乎决定了特定酶在像差诱导时的效率。 DNA修复抑制研究表明，RE诱导的双链断裂（DSB）修复在治疗后1小时内完成。使用胞嘧啶阿拉伯糖苷（araC）和蚜虫碱的G {dollar} sb1 {dollar}研究表明，在RE诱导的DSB修复中存在合成步骤。 3-氨基苯甲酰胺（3-AB），一种聚（ADP-核糖基）化抑制剂，可增强RE引起的G {dollar} sb1 {dollar}像差；它的作用似乎取决于RE诱导的DSB的数量或分布。在G {sb2 {dollar} RE处理期间，araC增强了染色单体的缺失和消色差病变的产量，但没有提高交换产量。 3-AB对G {sb2 {dollar}引起的像差影响很小。 DNA修复抑制实验表明，存在至少两种RE诱导的DSB修复机制：涉及合成步骤的重组修复，以及由聚（ADP-核糖基）化作用调控的连接修复。涉及不同对RE的相互作用实验为结扎修复提供了更多证据。然而，末端结构似乎影响了不同的RE诱导的DSB相互作用的能力。获得了有关开发两个不同系统以研究非随机染色体改变的初步数据。用不同的小鼠-仓鼠杂种细胞系进行的研究表明，X射线诱导了整个杂种基因组的像差。但是，丝裂霉素C优先在小鼠着丝粒异染色质中诱导畸变，后者富含重复的卫星DNA序列。不是I，一种具有罕见识别序列的酶，似乎仅在CHO核型中的少数位置诱导畸变。这两个系统可能为研究非随机染色体改变在细胞转化中的作用提供了有价值的工具。

著录项

  • 作者

    WINEGAR, RICHARD ALAN.;

  • 作者单位

    The University of Tennessee.;

  • 授予单位 The University of Tennessee.;
  • 学科 Biology Genetics.
  • 学位 Ph.D.
  • 年度 1987
  • 页码 160 p.
  • 总页数 160
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 遗传学;
  • 关键词

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