Modulation of restriction enzyme-induced damage by chemicals that interfere with cellular responses to DNA damage: a cytogenetic and pulsed-field gel analysis.
Modulation of restriction enzyme-induced damage by chemicals that interfere with cellular responses to DNA damage: a cytogenetic and pulsed-field gel analysis.
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通过干扰细胞对 DNA 损伤反应的化学物质来调节限制性内切酶诱导的损伤：细胞遗传学和脉冲场凝胶分析。
作者：
H. Chung;J. Phillips;R. Winegar;R. Preston;W. Morgan
限制性内切酶的电穿孔进入哺乳动物细胞会导致DNA双链断裂，从而导致染色体异常。研究了4种已知干扰细胞对DNA损伤反应的化学物质对AluI和Sau3AI诱发的染色体畸变率的影响，并用脉冲场凝胶电泳法测定了酶处理后不同时间DNA双链断裂的数目。聚(ADP-核糖)聚合酶抑制剂3-氨基苯甲酰胺(3AB)显着增加了交换和缺失的产率，并导致由酶诱导的双链断裂产量的微小但短暂的增加。1-β-D-阿拉伯呋喃葡萄糖胞嘧啶可通过直接作用于DNA聚合酶α和Delta或掺入DNA来抑制DNA修复，但其作用强度小于3AB，且不影响DNA双链断裂的数量。抑制聚合酶α和Delta的Aphidicolin对AluI诱发的畸变率没有影响，但增加了Sau3AI诱发的畸变率。复制后修复抑制剂咖啡因对任何一种酶诱导的畸变率都没有影响。用PFGE检测，APHIDICOIN和咖啡因都不能调节DNA双链断裂的数量。这些数据表明，多聚(ADP-核糖基)和聚合酶α和德尔塔是正常修复末端钝化或粘连的DNA双链断裂所需的细胞过程的重要组成部分。将这些数据与抑制剂对X射线诱导的像差频率的影响进行比较，我们得出的结论是，X射线诱导的像差可以由双链断裂的错接或不连接引起，特别是具有粘合末端的断裂，但这一过程只占诱导像差的一部分。
The electroporation of restriction enzymes into mammalian cells results in DNA double-strand breaks that can lead to chromosome aberrations. Four chemicals known to interfere with cellular responses to DNA damage were investigated for their effects on chromosome aberrations induced by AluI and Sau3AI; in addition, the number of DNA double-strand breaks at various times after enzyme treatment was determined by pulsed-field gel electrophoresis (PFGE). The poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide (3AB) dramatically increased the yield of exchanges and deletions and caused a small but transitory increase in the yield of double-strand breaks induced by the enzymes. 1-beta-D-Arabinofuranosylcytosine, which can inhibit DNA repair either by direct action on DNA polymerases alpha and delta or by incorporation into DNA, potentiated aberration induction but to a lesser extent than 3AB and did not affect the amount of DNA double-strand breakage. Aphidicolin, which inhibits polymerases alpha and delta, had no effect on AluI-induced aberrations but did increase the aberration yield induced by Sau3AI. The postreplication repair inhibitor caffeine had no effect on aberration yields induced by either enzyme. Neither aphidicolin nor caffeine modulated the amount of DNA double-strand breakage as measured by PFGE. These data implicate poly(ADP-ribosyl)ation and polymerases alpha and delta as important components of the cellular processes required for the normal repair of DNA double-strand breaks with blunt or cohesive ends. Comparison of these data with the effect of inhibitors on the frequency of X-ray-induced aberrations leads us to the conclusion that X-ray-induced aberrations can result from the misjoining or nonrejoining of double-strand breaks, particularly breaks with cohesive ends, but that this process accounts for only a portion of the induced aberrations.
通过电穿孔引入细胞后限制性核酸内切酶的细胞遗传学效应。
DOI：
--
发表时间：
1990
期刊：
Progress in clinical and biological research
影响因子：
--
作者：
Morgan,WF;Ager,D;Chung,HW;Ortiz,T;Phillips,JW;Winegar,RA
通讯作者：
Winegar,RA
通过脉冲场凝胶电泳分析中国仓鼠卵巢细胞中限制性内切酶诱导的 DNA 双链断裂：对染色体损伤的影响。
影响因子：
3.4
作者：
Ager,DD;Phillips,JW;Columna,EA;Winegar,RA;Morgan,WF
通讯作者：
Morgan,WF
细胞电穿孔是将限制性内切核酸酶引入细胞的高效方法。
DOI：
10.1016/0165-7992(89)90032-8
发表时间：
1989
期刊：
Mutation research
影响因子：
--
作者：
Winegar,RA;Phillips,JW;Youngblom,JH;Morgan,WF
通讯作者：
Morgan,WF