Sources and structures of mitotic crossovers that arise when BLM helicase is absent in Drosophila

• Published in: Genetics (Austin) Vol. 196; no. 1; pp. 107 - 118
• Main Authors: LaFave, Matthew C, Andersen, Sabrina L, Stoffregen, Eric P, Holsclaw, Julie K, Kohl, Kathryn P, Overton, Lewis J, Sekelsky, Jeff
• Format: Journal Article
• Language: English
• Published: United States Genetics Society of America 01.01.2014
• Subjects: Animals; Base Sequence; Cell division; Crossing Over, Genetic - drug effects; Crossing Over, Genetic - genetics; DNA Breaks, Double-Stranded; DNA End-Joining Repair - genetics; DNA Helicases - genetics; DNA Repair - genetics; Drosophila melanogaster; Drosophila melanogaster - genetics; Drosophila Proteins - genetics; Female; Genetics; Genomes; Hydroxyurea - pharmacology; Investigations; Male; Mitosis - genetics; Nucleic Acid Synthesis Inhibitors - pharmacology; Sequence Analysis, DNA; BLM helicase; crossovers; mitotic recombination
• ISSN: 1943-2631; 0016-6731; 1943-2631
• DOI: 10.1534/genetics.113.158618

The Bloom syndrome helicase, BLM, has numerous functions that prevent mitotic crossovers. We used unique features of Drosophila melanogaster to investigate origins and properties of mitotic crossovers that occur when BLM is absent. Induction of lesions that block replication forks increased crossover frequencies, consistent with functions for BLM in responding to fork blockage. In contrast, treatment with hydroxyurea, which stalls forks, did not elevate crossovers, even though mutants lacking BLM are sensitive to killing by this agent. To learn about sources of spontaneous recombination, we mapped mitotic crossovers in mutants lacking BLM. In the male germline, irradiation-induced crossovers were distributed randomly across the euchromatin, but spontaneous crossovers were nonrandom. We suggest that regions of the genome with a high frequency of mitotic crossovers may be analogous to common fragile sites in the human genome. Interestingly, in the male germline there is a paucity of crossovers in the interval that spans the pericentric heterochromatin, but in the female germline this interval is more prone to crossing over. Finally, our system allowed us to recover pairs of reciprocal crossover chromosomes. Sequencing of these revealed the existence of gene conversion tracts and did not provide any evidence for mutations associated with crossovers. These findings provide important new insights into sources and structures of mitotic crossovers and functions of BLM helicase.