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Aag fromBacillus subtilis has been implicated inin vitro removal of hypoxanthine and alkylated bases from DNA. The regulation of expression ofaag inB. subtilis and the resistance to genotoxic agents and mutagenic properties of an Aag-deficient strain were studied here. A strain with a transcriptionalaag-lacZ fusion expressed low levels of β-galactosidase during growth and early sporulation but exhibited increased transcription during late stages of this developmental process. Notably,aag-lacZ expression was higher inside the forespore than in the mother cell compartment, and this expression was abolished in asigG -deficient background, suggesting a forespore-specific mechanism ofaag transcription. Two additional findings supported this suggestion: (i) expression of anaag-yfp fusion was observed in the forespore, and (ii)in vivo mapping of theaag transcription start site revealed the existence of upstream regulatory sequences possessing homology to σG -dependent promoters. In comparison with the wild-type strain, disruption ofaag significantly reduced survival of sporulatingB. subtilis cells following nitrous acid or methyl methanesulfonate treatments, and the Rifr mutation frequency was significantly increased in anaag strain. These results suggest that Aag protects the genome of developingB. subtilis sporangia from the cytotoxic and genotoxic effects of base deamination and alkylation.IMPORTANCE In this study, evidence is presented revealing thataag , encoding a DNA glycosylase implicated in processing of hypoxanthine and alkylated DNA bases, exhibits a forespore-specific pattern of gene expression duringB. subtilis sporulation. Consistent with this spatiotemporal mode of expression, Aag was found to protect the sporulating cells of this microorganism from the noxious and mutagenic effects of base deamination and alkylation.

journal of bacteriology

Aag Hypoxanthine-DNA Glycosylase Is Synthesized in the Forespore Compartment and Involved in Counteracting the Genotoxic and Mutagenic Effects of Hypoxanthine and Alkylated Bases in DNA during Bacillus subtilis Sporulation