Viswanathan M, Lovett S T
Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02254-9110, USA.
Genetics. 1998 May;149(1):7-16. doi: 10.1093/genetics/149.1.7.
Mutations in the genes encoding single-strand DNA-specific exonucleases (ssExos) of Escherichia coli were examined for effects on mutation avoidance, UV repair, and conjugational recombination. Our results indicate complex and partially redundant roles for ssExos in these processes. Although biochemical experiments have implicated RecJ exonuclease, Exonuclease I (ExoI), and Exonuclease VII (ExoVII) in the methyl-directed mismatch repair pathway, the RecJ- ExoI- ExoVII- mutant did not exhibit a mutator phenotype in several assays for base substitution mutations. If these exonucleases do participate in mismatch excision, other exonucleases in E. coli can compensate for their loss. Frameshift mutations, however, were stimulated in the RecJ- ExoI- ExoVII- mutant. For acridine-induced frameshifts, this mutator effect was due to a synergistic effect of ExoI- and ExoVII- mutations, implicating both ExoI and ExoVII in avoidance of frameshift mutations. Although no single exonuclease mutant was especially sensitive to UV irradiation, the RecJ- ExoVII- double mutant was extremely sensitive. The addition of an ExoI- mutation augmented this sensitivity, suggesting that all three exonucleases play partially redundant roles in DNA repair. The ability to inherit genetic markers by conjugation was reduced modestly in the ExoI- RecJ- mutant, implying that the function of either ExoI or RecJ exonucleases enhances RecBCD-dependent homologous recombination.
我们检测了大肠杆菌中编码单链DNA特异性核酸外切酶(ssExos)的基因突变对突变避免、紫外线修复和接合重组的影响。我们的结果表明,ssExos在这些过程中发挥着复杂且部分冗余的作用。尽管生化实验表明RecJ核酸外切酶、核酸外切酶I(ExoI)和核酸外切酶VII(ExoVII)参与甲基导向的错配修复途径,但RecJ-ExoI-ExoVII-突变体在几种碱基替代突变检测中并未表现出突变体表型。如果这些核酸外切酶确实参与错配切除,那么大肠杆菌中的其他核酸外切酶可以弥补它们的缺失。然而,RecJ-ExoI-ExoVII-突变体中移码突变受到了刺激。对于吖啶诱导的移码突变,这种诱变效应是由于ExoI和ExoVII突变的协同作用,这表明ExoI和ExoVII都参与了移码突变的避免。尽管没有单个核酸外切酶突变体对紫外线照射特别敏感,但RecJ-ExoVII-双突变体对紫外线极其敏感。ExoI突变的加入增强了这种敏感性,这表明所有三种核酸外切酶在DNA修复中发挥着部分冗余的作用。ExoI-RecJ-突变体通过接合遗传标记的能力略有降低,这意味着ExoI或RecJ核酸外切酶的功能增强了RecBCD依赖性同源重组。