DNA end-joining driven by microhomologies catalyzed by nuclear extracts.
In a previous work we used an in vitro system for the generation and analysis of double-strand breaks (DSBs) using nuclear extracts from rat testes as a source of DSB activity. Since the recombination process can be triggered by the formation of DSB, in the present study we developed a strategy to isolate and characterize recombinant molecules using the same in vitro system. Our results indicate that the mechanism for the formation of recombinants was non-homologous end-joining driven by microhomologies. The procedure described here represents an alternative to investigate the mechanisms of DNA end-joining and other forms of DNA repair.
Boán
Francisco
F
Departamento de Bioquímica e Bioloxía Molecular, Facultade de Bioloxía, Universidade de Santiago de Compostela, E-15782 Santiago de Compostela, Spain.
Blanco
Miguel G
MG
Barros
Paula
P
Gómez-Márquez
Jaime
J
D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000737:chemistry*; D004260:DNA Repair / Q000235:genetics; D008297:Male; D009690:Nucleic Acid Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013737:Testis / Q000166:cytology
D011994:Recombinant Proteins; D004247:DNA