| Products/Services Used | Details | Operation |
|---|---|---|
| Codon Optimization | Expression and Purification of CtKu70/80 Codon-optimized synthetic DNA (Genscript, Piscataway, USA) encoding ctKu70 (N-terminal human rhinovirus 3C-protease cleav- able 6xHis-tag) and ctKu80 was PCR amplified and cloned into pACEBac1 and pIDC respectively and expressed using the MultiBac technology (Bieniossek et al. | Get A Quote |
DNA double-strand breaks (DSBs) threaten genome stability throughout life and are linked to tumorigenesis in humans. To initiate DSB repair by end joining or homologous recombination, the Mre11-nuclease Rad50-ATPase complex detects and processes diverse and obstructed DNA ends, but a structural mechanism is still lacking. Here we report cryo-EM structures of the E. coli Mre11-Rad50 homolog SbcCD in resting and DNA-bound cutting states. In the resting state, Mre11's nuclease is blocked by ATP-Rad50, and the Rad50 coiled coils appear flexible. Upon DNA binding, the two coiled coils zip up into a rod and, together with the Rad50 nucleotide-binding domains, form a clamp around dsDNA. Mre11 moves to the side of Rad... More
