Sharma R C, Smith K C
Department of Therapeutic Radiology, Stanford University School of Medicine, California 94305.
J Bacteriol. 1987 Oct;169(10):4559-64. doi: 10.1128/jb.169.10.4559-4564.1987.
Using strains of Escherichia coli K-12 that are deleted for the polA gene, we have reexamined the role of DNA polymerase I (encoded by polA) in postreplication repair after UV irradiation. The polA deletion (in contrast to the polA1 mutation) made uvrA cells very sensitive to UV radiation; the UV radiation sensitivity of a uvrA delta polA strain was about the same as that of a uvrA recF strain, a strain known to be grossly deficient in postreplication repair. The delta polA mutation interacted synergistically with a recF mutation in UV radiation sensitization, suggesting that the polA gene functions in pathways of postreplication repair that are largely independent of the recF gene. When compared to a uvrA strain, a uvrA delta polA strain was deficient in the repair of DNA daughter strand gaps, but not as deficient as a uvrA recF strain. Introduction of the delta polA mutation into uvrA recF cells made them deficient in the repair of DNA double-strand breaks after UV irradiation. The UV radiation sensitivity of a uvrA polA546(Ts) strain (defective in the 5'----3' exonuclease of DNA polymerase I) determined at the restrictive temperature was very close to that of a uvrA delta polA strain. These results suggest a major role for the 5'----3' exonuclease activity of DNA polymerase I in postreplication repair, in the repair of both DNA daughter strand gaps and double-strand breaks.
利用缺失polA基因的大肠杆菌K - 12菌株,我们重新研究了DNA聚合酶I(由polA编码)在紫外线照射后复制后修复中的作用。polA缺失(与polA1突变相反)使uvrA细胞对紫外线辐射非常敏感;uvrA ΔpolA菌株的紫外线辐射敏感性与uvrA recF菌株大致相同,uvrA recF菌株是一种已知在复制后修复方面严重缺陷的菌株。ΔpolA突变在紫外线辐射致敏方面与recF突变协同作用,表明polA基因在很大程度上独立于recF基因的复制后修复途径中发挥作用。与uvrA菌株相比,uvrA ΔpolA菌株在DNA子链缺口修复方面存在缺陷,但不如uvrA recF菌株缺陷严重。将ΔpolA突变引入uvrA recF细胞使其在紫外线照射后DNA双链断裂修复方面存在缺陷。在限制温度下测定的uvrA polA546(Ts)菌株(DNA聚合酶I的5'→3'外切核酸酶有缺陷)的紫外线辐射敏感性与uvrA ΔpolA菌株非常接近。这些结果表明DNA聚合酶I的5'→3'外切核酸酶活性在复制后修复、DNA子链缺口和双链断裂修复中起主要作用。