Role of Base Excision Repair (BER) in Transcription-associated Mutagenesis of Nutritionally Stressed Nongrowing Bacillus subtilis Cell Subpopulations
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Compelling evidence points to transcriptional processes as important factors contributing to stationary-phase associated mutagenesis. However, it has not been documented whether or not base excision repair mechanisms play a role in modulating mutagenesis under conditions of transcriptional derepression. Here, we report on a flow cytometry-based methodology that employs a fluorescent reporter system to measure at single-cell level, the occurrence of transcription-associated mutations in nutritionally stressed B. subtilis cultures. Using this approach, we demonstrate that (i) high levels of transcription correlates with augmented mutation frequency, and (ii) mutation frequency is enhanced in nongrowing population cells deficient for deaminated (Ung, YwqL) and oxidized guanine (GO) excision repair, strongly suggesting that accumulation of spontaneous DNA lesions enhance transcription-associated mutagenesis.
KeywordsBase Excision Repair Subtilis Cell Missense Allele Transcriptional Derepression Fluorescent Phenotype
This work was supported by the Consejo Nacional de Ciencia y Tecnología (CONACYT; Grants 205744 and 221231) of México and by the University of Guanajuato (Grants 936-2016 and 1090-2016). V. A–A was supported by a doctoral scholarship from CONACYT.
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Conflicts of Interest
The authors declare no conflict of interest.
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