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Publication Abstract from PubMed

Mitomycin C (MMC) repair factor A (mrfA) and factor B (mrfB), encode a conserved helicase and exonuclease that repair DNA damage in the soil-dwelling bacterium Bacillus subtilis. Here we have focused on the characterization of MrfB, a DEDDh exonuclease in the DnaQ superfamily. We solved the structure of the exonuclease core of MrfB to a resolution of 2.1 A, in what appears to be an inactive state. In this conformation, a predicted alpha-helix containing the catalytic DEDDh residue Asp172 adopts a random coil, which moves Asp172 away from the active site and results in the occupancy of only one of the two catalytic Mg2+ ions. We propose that MrfB resides in this inactive state until it interacts with DNA to become activated. By comparing our structure to an AlphaFold prediction as well as other DnaQ-family structures, we located residues hypothesized to be important for exonuclease function. Using exonuclease assays we show that MrfB is a Mg2+-dependent 3'-5' DNA exonuclease. We show that Leu113 aids in coordinating the 3' end of the DNA substrate, and that a basic loop is important for substrate binding. This work provides insight into the function of a recently discovered bacterial exonuclease important for the repair of MMC-induced DNA adducts.

Structural and biochemical characterization of the mitomycin C repair exonuclease MrfB.,Manthei KA, Munson LM, Nandakumar J, Simmons LA Nucleic Acids Res. 2024 Jun 24;52(11):6347-6359. doi: 10.1093/nar/gkae308. PMID:38661211^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.