Gamma-irradiation stimulates homology-directed DNA double-strand break repair in Drosophila embryo

Ducau, Judith; Bregliano, Jean-Claude

doi:10.1016/s0921-8777(00)00017-3

Cited by 11 publications

(6 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consistent with this, we have previously shown that a DSB caused by excision of a P element transposon in flies is readily repaired by a DNA ligase 4-independent end-joining process [17]. Interestingly, although Drosophila orthologs for the Pol X family DNA polymerases mu and lambda have not been identified [18], we and others have found evidence for polymerase activity in Drosophila end-joining repair [17], [19], [20]. Specifically, end joining in flies is often associated with the insertion of nucleotides at repair junctions, frequently involving imperfect repeats of 5–8 nucleotides.…”

Section: Introductionsupporting

confidence: 76%

Dual Roles for DNA Polymerase Theta in Alternative End-Joining Repair of Double-Strand Breaks in Drosophila

2010

View full text Add to dashboard Cite

DNA double-strand breaks are repaired by multiple mechanisms that are roughly grouped into the categories of homology-directed repair and non-homologous end joining. End-joining repair can be further classified as either classical non-homologous end joining, which requires DNA ligase 4, or “alternative” end joining, which does not. Alternative end joining has been associated with genomic deletions and translocations, but its molecular mechanism(s) are largely uncharacterized. Here, we report that Drosophila melanogaster DNA polymerase theta (pol theta), encoded by the mus308 gene and previously implicated in DNA interstrand crosslink repair, plays a crucial role in DNA ligase 4-independent alternative end joining. In the absence of pol theta, end joining is impaired and residual repair often creates large deletions flanking the break site. Analysis of break repair junctions from flies with mus308 separation-of-function alleles suggests that pol theta promotes the use of long microhomologies during alternative end joining and increases the likelihood of complex insertion events. Our results establish pol theta as a key protein in alternative end joining in Drosophila and suggest a potential mechanistic link between alternative end joining and interstrand crosslink repair.

show abstract

Section: Introductionsupporting

confidence: 76%

Dual Roles for DNA Polymerase Theta in Alternative End-Joining Repair of Double-Strand Breaks in Drosophila

2010

View full text Add to dashboard Cite

show abstract

“…Of the remaining 48 insertions, seven (14%) correspond to simple expansions of dinucleotides or small repeats flanking the insertions, and 41 (82%) have no match to the reference genome sequence and were thus classified as 'filler DNA' [13]. Filler DNA is a common outcome of the repair of DNA double-strand breaks by NHEJ in flies [33,34] and other organisms [35]. Filler DNA has been observed in several studies of DNA repair that use artificial DNA constructs where DNA double-strand breaks are induced and the products of the DNA repair can be recovered and sequenced.…”

Section: Resultsmentioning

confidence: 99%

“…Filler DNA has been observed in several studies of DNA repair that use artificial DNA constructs where DNA double-strand breaks are induced and the products of the DNA repair can be recovered and sequenced. In most cases, only a few nucleotides (or none) are added to the repaired junctions, but in some instances large insertions are created [13,33,35]. …”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Mutation spectrum of Drosophila CNVs revealed by breakpoint sequencing

2012

View full text Add to dashboard Cite

BackgroundThe detailed study of breakpoints associated with copy number variants (CNVs) can elucidate the mutational mechanisms that generate them and the comparison of breakpoints across species can highlight differences in genomic architecture that may lead to lineage-specific differences in patterns of CNVs. Here, we provide a detailed analysis of Drosophila CNV breakpoints and contrast it with similar analyses recently carried out for the human genome.ResultsBy applying split-read methods to a total of 10x coverage of 454 shotgun sequence across nine lines of D. melanogaster and by re-examining a previously published dataset of CNVs detected using tiling arrays, we identified the precise breakpoints of more than 600 insertions, deletions, and duplications. Contrasting these CNVs with those found in humans showed that in both taxa CNV breakpoints fall into three classes: blunt breakpoints; simple breakpoints associated with microhomology; and breakpoints with additional nucleotides inserted/deleted and no microhomology. In both taxa CNV breakpoints are enriched with non-B DNA sequence structures, which may impair DNA replication and/or repair. However, in contrast to human genomes, non-allelic homologous-recombination (NAHR) plays a negligible role in CNV formation in Drosophila. In flies, non-homologous repair mechanisms are responsible for simple, recurrent, and complex CNVs, including insertions of de novo sequence as large as 60 bp.ConclusionsHumans and Drosophila differ considerably in the importance of homology-based mechanisms for the formation of CNVs, likely as a consequence of the differences in the abundance and distribution of both segmental duplications and transposable elements between the two genomes.

show abstract

“…Some researchers have done "in vivo extract" studies by injecting repair substrates into embryos, then recovering the products for molecular analysis. Examples include repair of DSBs after excision of a P element from a plasmid (O'Brochta et al 1991;Beall and Rio 1996), or after introduction of linear plasmid and repair template (Ducau et al 2000).…”

Section: Other Assaysmentioning

confidence: 99%

DNA Repair inDrosophila: Mutagens, Models, and Missing Genes

2017

View full text Add to dashboard Cite

The numerous processes that damage DNA are counterbalanced by a complex network of repair pathways that, collectively, can mend diverse types of damage. Insights into these pathways have come from studies in many different organisms, including Drosophila melanogaster. Indeed, the first ideas about chromosome and gene repair grew out of Drosophila research on the properties of mutations produced by ionizing radiation and mustard gas. Numerous methods have been developed to take advantage of Drosophila genetic tools to elucidate repair processes in whole animals, organs, tissues, and cells. These studies have led to the discovery of key DNA repair pathways, including synthesis-dependent strand annealing, and DNA polymerase theta-mediated end joining. Drosophila appear to utilize other major repair pathways as well, such as base excision repair, nucleotide excision repair, mismatch repair, and interstrand crosslink repair. In a surprising number of cases, however, DNA repair genes whose products play important roles in these pathways in other organisms are missing from the Drosophila genome, raising interesting questions for continued investigations.

show abstract

Gamma-irradiation stimulates homology-directed DNA double-strand break repair in Drosophila embryo

Cited by 11 publications

References 44 publications

Dual Roles for DNA Polymerase Theta in Alternative End-Joining Repair of Double-Strand Breaks in Drosophila

Dual Roles for DNA Polymerase Theta in Alternative End-Joining Repair of Double-Strand Breaks in Drosophila

Mutation spectrum of Drosophila CNVs revealed by breakpoint sequencing

DNA Repair inDrosophila: Mutagens, Models, and Missing Genes

Contact Info

Product

Resources

About