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Genetics, Vol. 168, 2067-2076, December 2004, Copyright © 2004
doi:10.1534/genetics.104.033902

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End-Joining Repair of Double-Strand Breaks in Drosophila melanogaster Is Largely DNA Ligase IV Independent

Mitch McVey*,{dagger},1, Dora Radut* and Jeff J. Sekelsky*,{ddagger}

* Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599
{dagger} SPIRE Program, University of North Carolina, Chapel Hill, North Carolina 27599
{ddagger} Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, North Carolina 27599

1 Corresponding author: Department of Biology, CB 3280, 303 Fordham Hall, University of North Carolina, Chapel Hill, NC 27599-3280.
E-mail: mmcvey{at}email.unc.edu

Repair of DNA double-strand breaks can occur by either nonhomologous end joining or homologous recombination. Most nonhomologous end joining requires a specialized ligase, DNA ligase IV (Lig4). In Drosophila melanogaster, double-strand breaks created by excision of a P element are usually repaired by a homologous recombination pathway called synthesis-dependent strand annealing (SDSA). SDSA requires strand invasion mediated by DmRad51, the product of the spn-A gene. In spn-A mutants, repair proceeds through a nonconservative pathway involving the annealing of microhomologies found within the 17-nt overhangs produced by P excision. We report here that end joining of P-element breaks in the absence of DmRad51 does not require Drosophila LIG4. In wild-type flies, SDSA is sometimes incomplete, and repair is finished by an end-joining pathway that also appears to be independent of LIG4. Loss of LIG4 does not increase sensitivity to ionizing radiation in late-stage larvae, but lig4 spn-A double mutants do show heightened sensitivity relative to spn-A single mutants. Together, our results suggest that a LIG4-independent end-joining pathway is responsible for the majority of double-strand break repair in the absence of homologous recombination in flies.




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