MFD PROMOTES MUTAGENESIS AND PREVENTS PROTEIN DAMAGE IN CELLS EXPERIENCING OXIDATIVE STRESS IN BACILLUS SUBTILIS

Summary This application explores the functions of a DNA repair factor as a new element affecting evolutionand adaptation of bacteria, including those that cause infections. This is of scientific interest because itinvolves the discovery of molecular pathways that affect bacterial cell physiology and evolution inconditions of stress. Thus, the knowledge generated by this application has the potential to uncover noveltargets for the treatment of pernicious bacterial infections. The hypothesis under study is that the Mfdfactor, previously studied in the context of high-fidelity DNA repair of transcribed regions, mitigates thetoxic effects associated with oxidative stress by mediating error-prone DNA repair of DNA lesions andactivating expression of genes that prevent cell death. This hypothesis will be tested using genetic,biochemistry, and systems biology approaches. In Aim 1, we will test the concept that Mfd-dependentmutagenesis operates at genes that are highly transcribed in cells experiencing oxidative stress. Thisconcept will be examined innovatively by measuring mutagenesis at the genomic level and in the absenceof selection. Aim 2 will elucidate whether Mfd is important for activating expression of factors thatprotect against protein damage caused by oxidative stress independently of its previously associatedDNA repair functions. In conclusion, the experiments in this application are straight-forward and self-contained and seek to determine new functions of the Mfd factor. Mfd is well-conserved in bacterialpathogens and so the research proposed can potentially uncover a novel therapeutic target.