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Originally published as Genetics Published Articles Ahead of Print on September 2, 2005.
Genetics, Vol. 171, 1535-1548, December 2005, Copyright © 2005
doi:10.1534/genetics.105.046144
Capture of Extranuclear DNA at Fission Yeast Double-Strand Breaks
Anabelle Decottignies1
Cellular Genetics, Christian de Duve Institute of Cellular Pathology, Catholic University of Louvain, 1200 Brussels, Belgium
1 Address for correspondence: Cellular Genetics, Christian de Duve Institute of Cellular Pathology, Catholic University of Louvain, Ave. Hippocrate 74+3, 1200 Brussels, Belgium.
E-mail: anabelle.decottignies{at}gece.ucl.ac.be
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.
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