Antibiotic resistance in Bacillus subtilis as affected by transcriptional derepression and the stringent response

Bacterial cells under conditions of starvation or prolonged non-lethal selective pressures accumulate mutations in highly transcribed genes. This process is part of cellular programs to increase genetic diversity in conditions of stress, also known as stationary phase or stress-induced mutagenesis. This experiment investigated mutation frequencies for antibiotic resistance as affected by the stringent response. The stringent response is a global cellular process that initiates at the cessation of growth and mediates changes in gene expression that repress synthesis of ribosome components. We used Bacillus subtilis strains that differ in RelA proficiency. The relA gene controls the synthesis of (p)ppGpp, the signaling molecule which mediates the stringent response. Since genes involved in protein synthesis are repressed during the stringent response, we hypothesize that relaxed mutants express a higher accumulation of mutations that confer resistance to tetracycline than cells that become stringent. Resistance to tetracycline may be acquired by altering components of the small subunit of bacterial ribosomes. Utilizing an overlay procedure and increasing times of incubation under nutritional stress, stationary cells were prompted for resistance to tetracycline. Our results showed that relA- cells expressed a higher accumulation of Tcr mutations than the one observed in wild type cells. These results provide evidence that transcriptional derepression in cells under non-lethal stress mediates mutagenic events. Implications in antibiotic resistance are further discussed