Wang Z, Wu X, Friedberg E C
Department of Pathology, University of Texas Southwestern Medical Center, Dallas 75235-9072.
Mol Cell Biol. 1993 Feb;13(2):1051-8. doi: 10.1128/mcb.13.2.1051-1058.1993.
Base excision repair is an important mechanism for correcting DNA damage produced by many physical and chemical agents. We have examined the effects of the REV3 gene and the DNA polymerase genes POL1, POL2, and POL3 of Saccharomyces cerevisiae on DNA repair synthesis is nuclear extracts. Deletional inactivation of REV3 did not affect repair synthesis in the base excision repair pathway. Repair synthesis in nuclear extracts of pol1, pol2, and pol3 temperature-sensitive mutants was normal at permissive temperatures. However, repair synthesis in pol2 nuclear extracts was defective at the restrictive temperature of 37 degrees C and could be complemented by the addition of purified yeast DNA polymerase epsilon. Repair synthesis in pol1 nuclear extracts was proficient at the restrictive temperature unless DNA polymerase alpha was inactivated prior to the initiation of DNA repair. Thermal inactivation of DNA polymerase delta in pol3 nuclear extracts enhanced DNA repair synthesis approximately 2-fold, an effect which could be specifically reversed by the addition of purified yeast DNA polymerase delta to the extract. These results demonstrate that DNA repair synthesis in the yeast base excision repair pathway is catalyzed by DNA polymerase epsilon but is apparently modulated by the presence of DNA polymerases alpha and delta.
碱基切除修复是纠正由多种物理和化学因子产生的DNA损伤的重要机制。我们研究了酿酒酵母的REV3基因以及DNA聚合酶基因POL1、POL2和POL3对核提取物中DNA修复合成的影响。REV3的缺失失活不影响碱基切除修复途径中的修复合成。pol1、pol2和pol3温度敏感突变体的核提取物中的修复合成在允许温度下是正常的。然而,pol2核提取物中的修复合成在37℃的限制温度下有缺陷,并且可以通过添加纯化的酵母DNA聚合酶ε来补充。pol1核提取物中的修复合成在限制温度下是有效的,除非在DNA修复开始之前DNA聚合酶α被灭活。pol3核提取物中DNA聚合酶δ的热失活使DNA修复合成增强了约2倍,通过向提取物中添加纯化的酵母DNA聚合酶δ可以特异性地逆转这种效应。这些结果表明,酵母碱基切除修复途径中的DNA修复合成由DNA聚合酶ε催化,但显然受到DNA聚合酶α和δ的存在的调节。