Rad51-mediated replication of damaged templates relies on monoSUMOylated DDK kinase

• Published in: Nature communications 2022-05, Vol.13 (1), p.2480-2480, Article 2480
• Main Authors: Joseph, Chinnu Rose, Dusi, Sabrina, Giannattasio, Michele, Branzei, Dana
• Format: Article
• Language: English
• Subjects: 101/28; 38/1; 38/15; 38/22; 38/23; 38/71; 38/77; 42/41; 45; 45/15; 45/22; 631/337/1427/2190; 631/337/149; 631/337/151/2356; 82/1; 82/29; Damage tolerance; DDT; Deoxyribonucleic acid; DNA; DNA Damage; DNA Repair; DNA Replication; Genomes; Genotoxicity; Homologous Recombination; Homology; Humanities and Social Sciences; Kinases; multidisciplinary; Mutation; Phenotypes; Rad51 Recombinase - genetics; Rad51 Recombinase - metabolism; Recombinase; Replication; Replication forks; Replication initiation; Science; Science (multidisciplinary); Single-stranded DNA
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-30215-9

DNA damage tolerance (DDT), activated by replication stress during genome replication, is mediated by translesion synthesis and homologous recombination (HR). Here we uncover that DDK kinase, essential for replication initiation, is critical for replication-associated recombination-mediated DDT. DDK relies on its multi-monoSUMOylation to facilitate HR-mediated DDT and optimal retention of Rad51 recombinase at replication damage sites. Impairment of DDK kinase activity, reduced monoSUMOylation and mutations in the putative SUMO Interacting Motifs (SIMs) of Rad51 impair replication-associated recombination and cause fork uncoupling with accumulation of large single-stranded DNA regions at fork branching points. Notably, genetic activation of salvage recombination rescues the uncoupled fork phenotype but not the recombination-dependent gap-filling defect of DDK mutants, revealing that the salvage recombination pathway operates preferentially proximal to fork junctions at stalled replication forks. Overall, we uncover that monoSUMOylated DDK acts with Rad51 in an axis that prevents replication fork uncoupling and mediates recombination-dependent gap-filling. Joseph et al. reveal that monoSUMOylated DDK kinase, implicated in replication initiation, acts with Rad51 recombinase to prevent replication fork uncoupling and to mediate recombination-dependent gap-filling in the presence of genotoxic stress.