Lig4 and Rad54 are required for repair of DNA double-strand breaks induced by P-element excision in Drosophila

Ron J. Romeijn, Marcin M. Gorski, Martijn A. Van Schie, Jasprina N. Noordermeer, Leon H. Mullenders, Wouter Ferro, Albert Pastink

Research output: Contribution to journalArticle

Abstract

Site-specific double-strand breaks (DSBs) were generated in the white gene located on the X chromosome of Drosophila by excision of the w hd P-element. To investigate the role of nonhomologous end joining (NHEJ) and homologous recombination (HR) in the repair of these breaks, the w wd P-element was mobilized in flies carrying mutant alleles of either lig4 or rad54. The survival of both lig4- and rad54-deficient males was reduced to 25% in comparison to the wild type, indicating that both NHEJ and HR are involved in the repair P-induced gaps in males. Survival of lig4-deficient females was not affected at all, implying that HR using the homologous chromosome as a template can partially compensate for the impaired NHEJ pathway. In rad54 mutant females survival was reduced to 70% after w hd excision. PCR analysis indicated that the undamaged homologous chromosome may compensate for the potential loss of the broken chromosome in rad54 mutant females after excision. Molecular analysis of the repair junctions revealed microhomology (2-8 bp)-dependent DSB repair in most products. In the absence of Lig4, the 8-bp target site duplication is used more frequently for repair. Our data indicate the presence of efficient alternative end-joining mechanisms, which partly depend on the presence of microhomology but do not require Lig4.

Original languageEnglish
Pages (from-to)795-806
Number of pages12
JournalGenetics
Volume169
Issue number2
DOIs
Publication statusPublished - Feb 2005

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Double-Stranded DNA Breaks
Drosophila
Chromosomes
Homologous Recombination
Recombinational DNA Repair
X Chromosome
Diptera
Alleles
Polymerase Chain Reaction
Genes

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Romeijn, R. J., Gorski, M. M., Van Schie, M. A., Noordermeer, J. N., Mullenders, L. H., Ferro, W., & Pastink, A. (2005). Lig4 and Rad54 are required for repair of DNA double-strand breaks induced by P-element excision in Drosophila. Genetics, 169(2), 795-806. https://doi.org/10.1534/genetics.104.033464

Lig4 and Rad54 are required for repair of DNA double-strand breaks induced by P-element excision in Drosophila. / Romeijn, Ron J.; Gorski, Marcin M.; Van Schie, Martijn A.; Noordermeer, Jasprina N.; Mullenders, Leon H.; Ferro, Wouter; Pastink, Albert.

In: Genetics, Vol. 169, No. 2, 02.2005, p. 795-806.

Research output: Contribution to journalArticle

Romeijn, RJ, Gorski, MM, Van Schie, MA, Noordermeer, JN, Mullenders, LH, Ferro, W & Pastink, A 2005, 'Lig4 and Rad54 are required for repair of DNA double-strand breaks induced by P-element excision in Drosophila', Genetics, vol. 169, no. 2, pp. 795-806. https://doi.org/10.1534/genetics.104.033464
Romeijn RJ, Gorski MM, Van Schie MA, Noordermeer JN, Mullenders LH, Ferro W et al. Lig4 and Rad54 are required for repair of DNA double-strand breaks induced by P-element excision in Drosophila. Genetics. 2005 Feb;169(2):795-806. https://doi.org/10.1534/genetics.104.033464
Romeijn, Ron J. ; Gorski, Marcin M. ; Van Schie, Martijn A. ; Noordermeer, Jasprina N. ; Mullenders, Leon H. ; Ferro, Wouter ; Pastink, Albert. / Lig4 and Rad54 are required for repair of DNA double-strand breaks induced by P-element excision in Drosophila. In: Genetics. 2005 ; Vol. 169, No. 2. pp. 795-806.
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