Hagensee M E, Timme T L, Bryan S K, Moses R E
Proc Natl Acad Sci U S A. 1987 Jun;84(12):4195-9. doi: 10.1073/pnas.84.12.4195.
Strains of Escherichia coli possessing the pcbA1 mutation, a functional DNA polymerase I, and a temperature-sensitive mutation in DNA polymerase III can survive at the restrictive temperature (43 degrees C) for DNA polymerase III. The mutation rate of the bacterial genome of such strains after exposure to either UV light or ethyl methanesulfonate was measured by its rifampicin resistance or amino acid requirements. In addition, Weigle mutagenesis of preirradiated lambda phage was also measured. In all cases, no increase in mutagenesis was noted at the restrictive temperature for DNA polymerase III. Introduction of a cloned DNA polymerase III gene returned the mutation rate of the bacterial genome as well as the Weigle mutagenesis to normal at 43 degrees C. Using a recA-lacZ fusion, the SOS response after UV irradiation was measured and found to be normal at the restrictive and permissive temperature for DNA polymerase III, as was induction of lambda prophage. Recombination was also normal at either temperature. Our studies demonstrate that a functional DNA polymerase III is strictly required for mutagenesis at a step other than SOS induction.
具有pcbA1突变、功能性DNA聚合酶I以及DNA聚合酶III温度敏感突变的大肠杆菌菌株,能够在DNA聚合酶III的限制温度(43摄氏度)下存活。通过其对利福平的抗性或氨基酸需求,来测定此类菌株的细菌基因组在暴露于紫外线或甲磺酸乙酯后产生的突变率。此外,还测定了预照射的λ噬菌体的韦格勒诱变情况。在所有情况下,在DNA聚合酶III的限制温度下均未观察到诱变增加。导入克隆的DNA聚合酶III基因后,细菌基因组的突变率以及韦格勒诱变在43摄氏度时恢复正常。使用recA-lacZ融合蛋白,测定了紫外线照射后的SOS反应,发现在DNA聚合酶III的限制温度和允许温度下该反应均正常,λ原噬菌体的诱导情况也是如此。在这两个温度下重组也均正常。我们的研究表明,在SOS诱导以外的某个步骤进行诱变时,严格需要功能性DNA聚合酶III。