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bioRxiv · 10.1101/2020.11.24.395657

Deuterium-depletion has no significant impact on the mutation rate of Escherichia coli, invalidating the Double D-Bridge Hypothesis

Ajibola, W.·Karcagi, I.·Somlyai, G.·Somlyai, I.·Tamas, F.
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Abstract

Deuterium (D), the second most abundant isotope of hydrogen is present in natural waters at an approximate concentration of 145-155 ppm (cca. 1.5E-4 atom/atom). D is known to influence various biological processes due to its physical and chemical properties, which significantly differ from those of hydrogen. For example, increasing D-concentration to >1000-fold above its natural abundance has been shown to increase the frequency of genetic mutations in several species. The Double D-Bridge Hypothesis, formulated with the intent of explaining the mechanism of D-mutagenicity is based on the calculation that the theoretical frequency of A:T base pairs that comprise two D-bridges instead of H-bridges is 2E-8, which is equal to the mutation rate of certain species. To experimentally challenge this hypothesis, and to infer the mutagenicity of D present at natural concentrations, we investigated the effect of a nearly 100-fold reduction of D concentration on the bacterial mutation rate. Using fluctuation tests, we measured the mutation rate of three Escherichia coli genes (cycA, ackA and galK) in media containing D at either <2 ppm or 150 ppm concentrations. Out of 15 pair-wise fluctuation analyses, nine indicated a significant decrease, while three marked the significant increase of the mutation/culture value upon D-depletion. Overall, growth in D-depleted minimal medium led to a geometric mean of 0.663-fold (95% confidence interval: 0.483-0.911) change in the mutation rate. This falls nowhere near the expected 10,000-fold reduction, indicating that the Double D-Bridge Hypothesis does not hold. In addition, the combined results did not display a statistically significant change in the mutation/culture value, the mutation rate or the mutant frequency upon D-depletion. The potential mutagenic effect of D present at natural concentrations on E. coli is therefore below the limit of detection using the indicated methods.

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Ajibola, W., Karcagi, I., Somlyai, G., Somlyai, I., Tamas, F.. 2020-11-24. Deuterium-depletion has no significant impact on the mutation rate of Escherichia coli, invalidating the Double D-Bridge Hypothesis. https://doi.org/10.1101/2020.11.24.395657

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