Budke Brian, Kuzminov Andrei
B103 C&LSL, 601 South Goodwin Ave., Urbana, IL 61801-3709, USA.
J Bacteriol. 2006 Sep;188(18):6553-60. doi: 10.1128/JB.00447-06.
Endonuclease V, encoded by the nfi gene, initiates removal of the base analogs hypoxanthine and xanthine from DNA, acting to prevent mutagenesis from purine base deamination within the DNA. On the other hand, the RdgB nucleotide hydrolase in Escherichia coli is proposed to prevent hypoxanthine and xanthine incorporation into DNA by intercepting the noncanonical DNA precursors dITP and dXTP. Because many base analogs are mutagenic when incorporated into DNA, it is intuitive to think of RdgB as acting to prevent similar mutagenesis from deaminated purines in the DNA precursor pools. To test this idea, we used a set of Claire Cupples' strains to detect changes in spontaneous mutagenesis spectra, as well as in nitrous acid-induced mutagenesis spectra, in wild-type cells and in rdgB single, nfi single, and rdgB nfi double mutants. We found neither a significant increase in spontaneous mutagenesis in rdgB and nfi single mutants or the double mutant nor any changes in nitrous acid-induced mutagenesis for rdgB mutant strains. We conclude that incorporation of deaminated purines into DNA is nonmutagenic.
由nfi基因编码的核酸内切酶V启动从DNA中去除碱基类似物次黄嘌呤和黄嘌呤的过程,起到防止DNA内嘌呤碱基脱氨基引起诱变的作用。另一方面,有人提出大肠杆菌中的RdgB核苷酸水解酶通过拦截非经典DNA前体dITP和dXTP来防止次黄嘌呤和黄嘌呤掺入DNA。由于许多碱基类似物掺入DNA时具有诱变性,因此直观地认为RdgB的作用是防止DNA前体库中脱氨基嘌呤产生类似的诱变作用。为了验证这一想法,我们使用了一组克莱尔·卡普尔斯菌株来检测野生型细胞以及rdgB单突变体、nfi单突变体和rdgB nfi双突变体中自发诱变谱以及亚硝酸诱导诱变谱的变化。我们发现rdgB和nfi单突变体或双突变体的自发诱变均未显著增加,并且rdgB突变菌株的亚硝酸诱导诱变也没有任何变化。我们得出结论,脱氨基嘌呤掺入DNA不会产生诱变作用。