Abstract
The ability of cells to maintain their genomic integrity and accurately repair any DNA damage is crucial to their continued survival and functioning. One such type of damage, double-stranded breaks, is primarily addressed in Escherichia coli (E. coli) by RecA-mediated homologous recombination, a highly conserved pathway in which the damaged region is matched to a homology, which is then used as a template to bridge the gap. This strand exchange event forms a secondary structure known as a Holliday junction, which must be resolved before the gap can be fully repaired. Although the broad outline of this pathway is well understood, there are still many details that require further investigation. In this study, we examined two proteins, RadA and RadD, which have both been broadly implicated in double-stranded break repair. Previous results have shown shared functions and shared mutant phenotypes, but have not conclusively shown that they physically interact. To that end, we used a yeast two-hybrid assay, which showed an interaction, followed by further yeast two-hybrids in order to examine whether specific RadA mutants could disrupt this interaction.