Roles for mycobacterial DinB2 in frameshift and substitution mutagenesis

• DOI: 10.7554/elife.83094
• Publication Date: 2023-05-04T17:00:23Z
• Authors: Pierre Dupuy; Shreya Ghosh; Allison Fay; Oyindamola Adefisayo; Richa Gupta; Stewart Shuman; Michael S Glickman
• Keywords: Microbiology and Infectious Disease; DNA Repair; QH301-705.5; Science; Mycobacterium smegmatis; Q; R; Mycobacterium tuberculosis; DNA damage response; 3. Good health; [SDV] Life Sciences [q-bio]; Bacterial Proteins; Mutagenesis; Medicine; antimicrobial resistance; Biology (General); Frameshift Mutation; Mycobacterium tuberculoiss; mutagenesis

Translesion synthesis by translesion polymerases is a conserved mechanism of DNA damage tolerance. In bacteria, DinB enzymes are the widely distributed promutagenic translesion polymerases. The role of DinBs in mycobacterial mutagenesis was unclear until recent studies revealed a role for mycobacterial DinB1 in substitution and frameshift mutagenesis, overlapping with that of translesion polymerase DnaE2. Mycobacterium smegmatis encodes two additional DinBs (DinB2 and DinB3) and Mycobacterium tuberculosis encodes DinB2, but the roles of these polymerases in mycobacterial damage tolerance and mutagenesis is unknown. The biochemical properties of DinB2, including facile utilization of ribonucleotides and 8-oxo-guanine, suggest that DinB2 could be a promutagenic polymerase. Here, we examine the effects of DinB2 and DinB3 overexpression in mycobacterial cells. We demonstrate that DinB2 can drive diverse substitution mutations conferring antibiotic resistance. DinB2 induces frameshift mutations in homopolymeric sequences, both in vitro and in vivo. DinB2 switches from less to more mutagenic in the presence of manganese in vitro. This study indicates that DinB2 may contribute to mycobacterial mutagenesis and antibiotic resistance acquisition in combination with DinB1 and DnaE2.