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Genetics. 2005 Dec;171(4):1535-48. Epub 2005 Sep 2.

Capture of extranuclear DNA at fission yeast double-strand breaks.

Author information

1
Cellular Genetics, Christian de Duve Institute of Cellular Pathology, Catholic University of Louvain, Avenue Hippocrate 74+3, 1200 Brussels, Belgium. anabelle.decottignies@gece.ucl.ac.be

Abstract

Proper repair of DNA double-strand breaks (DSBs) is necessary for the maintenance of genomic integrity. Here, a new simple assay was used to study extrachromosomal DSB repair in Schizosaccharomyces pombe. Strikingly, DSB repair was associated with the capture of fission yeast mitochondrial DNA (mtDNA) at high frequency. Capture of mtDNA fragments required the Lig4p/Pku70p nonhomologous end-joining (NHEJ) machinery and its frequency was highly increased in fission yeast cells grown to stationary phase. The fission yeast Mre11 complex Rad32p/Rad50p/Nbs1p was also required for efficient capture of mtDNA at DSBs, supporting a role for the complex in promoting intermolecular ligation. Competition assays further revealed that microsatellite DNA from higher eukaryotes was preferentially captured at yeast DSBs. Finally, cotransformation experiments indicated that, in NHEJ-deficient cells, capture of extranuclear DNA at DSBs was observed if homologies--as short as 8 bp--were present between DNA substrate and DSB ends. Hence, whether driven by NHEJ, microhomology-mediated end-joining, or homologous recombination, DNA capture associated with DSB repair is a mutagenic process threatening genomic stability.

PMID:
16143617
PMCID:
PMC1456082
DOI:
10.1534/genetics.105.046144
[Indexed for MEDLINE]
Free PMC Article

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