dUTPase modulates mycobacterial homologous recombination and interacts with the AdnAB helicase–nuclease

Source: PubMed Central Open Access, NCBI / U.S. National Library of Medicine

Abstract This study identifies a previously unrecognized interaction betweendUTPase (Dut) and the AdnAB homologous recombination complex. Using a combination of yeast two-hybrid screening, mycobacterial protein fragment complementation, and biochemical analyses with purified proteins, we show that dUTPase physically interacts with the N-terminal region of AdnA and modulates the activity of the AdnAB helicase–nuclease complex. Biochemical assays demonstrate that Dut enhances AdnAB activity on DNA substrates and alters the AdnAB–DNA interaction. Mutational perturbation of Dut, including catalytic inactivation or deletion of a mycobacteria-specific surface loop, reduces its stimulatory effect on AdnABand decreases recombination efficiency in mycobacterial cells. Together, these results support a functional connection between dUTPase and the AdnAB DNA-processing machinery and suggest a potential link between nucleotide metabolism and DNA repair pathways. Graphical Abstract Graphical Abstract For image description, please refer to the figure legend and surrounding text. http://www.w3.org/1999/xlink float portrait gkag660gra.jpg float ga1 portrait graphical