The Role of Blm Helicase in Homologous Recombination, Gene Conversion Tract Length, and Recombination Between Diverged Sequences in Drosophila melanogaster

• Published in: Genetics (Austin) 2017-11, Vol.207 (3), p.923-933
• Main Authors: Ertl, Henry A, Russo, Daniel P, Srivastava, Noori, Brooks, Joseph T, Dao, Thu N, LaRocque, Jeannine R
• Format: Article
• Language: English
• Subjects: Animals; Annealing; ATP-Binding Cassette Transporters - genetics; Cell cycle; Conversion; Crossovers; Deoxyribonucleic acid; DNA; DNA damage; DNA End-Joining Repair; DNA helicase; DNA Helicases - genetics; DNA Helicases - metabolism; DNA repair; Drosophila; Drosophila melanogaster; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Eye Proteins - genetics; Gene Conversion; Gene sequencing; Genetic recombination; Genetics; Genomes; Heterozygotes; Homologous recombination; Homology; Insects; Investigations; Loss of Function Mutation; Mutants; Mutation; Non-homologous end joining; Nucleotide sequence; Recombinational DNA Repair; Repair; Saccharomyces cerevisiae; Yeast
• ISSN: 1943-2631; 0016-6731; 1943-2631
• DOI: 10.1534/genetics.117.300285

DNA double-strand breaks (DSBs) are a particularly deleterious class of DNA damage that threatens genome integrity. DSBs are repaired by three pathways: nonhomologous-end joining (NHEJ), homologous recombination (HR), and single-strand annealing (SSA). ( ) is the ortholog of and human , and has been shown to suppress crossovers in mitotic cells and repair mitotic DNA gaps via HR. To further elucidate the role of DmBlm in repair of a simple DSB, and in particular recombination mechanisms, we utilized the Direct Repeat of (DR- ) and Direct Repeat of (DR- ) repair assays in multiple mutant allele backgrounds. null and helicase-dead mutants both demonstrated a decrease in repair by noncrossover HR, and a concurrent increase in non-HR events, possibly including SSA, crossovers, deletions, and NHEJ, although detectable processing of the ends was not significantly impacted. Interestingly, gene conversion tract lengths of HR repair events were substantially shorter in null but not helicase-dead mutants, compared to heterozygote controls. Using DR- , we found that, in contrast to Sgs1, DmBlm is not required for suppression of recombination between diverged sequences. Taken together, our data suggest that DmBlm helicase function plays a role in HR, and the steps that contribute to determining gene conversion tract length are helicase-independent.