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. 2017 Jul 20;7:6043. doi: 10.1038/s41598-017-06206-y

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The effect of dut and dcd:dut mutations on the mutation rate, DNA uracilation and gene expression. (A) Mutation rates of mutant M. smegmatis strains. Means ± SE are calculated from (3*3) independent colonies from each mutation. Note that mutation rates of dut mutant strains directly correlate with the in vitro activity-loss of the corresponding mutant enzyme. Significance levels: P = 0.000115 for S148A, P = 0.000034 for T138stop and P =0.000004 for D83N. “+” denotes the wt enzyme. (B) Genomic uracil content of the mutant strains compared to the wt strain. Uracil contents were calculated from three independent strains from each mutant and normalized to the wt strain; means ± SE are plotted. Note that the genomic uracil content of dut mutant strains directly correlates with the in vitro activity-loss of the corresponding mutant enzyme. Significance levels: P = 0.074 for T138stop, P = 0.018 for D83N. (C) Quantitation of dut and dcd:dut expression levels in the wt and mutant M. smegmatis strains. mRNA levels were calculated from three independent strains from each mutant and normalized to their respective wt strain; means ± SE are plotted. Significance level: P = 0.029 for the A115F mutant.