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TitleMechanism of DNA End Sensing and Processing by the Mre11-Rad50 Complex.
Journal, issue, pagesMol Cell, Vol. 76, Issue 3, Page 382-394.e6, Year 2019
Publish dateNov 7, 2019
AuthorsLisa Käshammer / Jan-Hinnerk Saathoff / Katja Lammens / Fabian Gut / Joseph Bartho / Aaron Alt / Brigitte Kessler / Karl-Peter Hopfner /
PubMed AbstractDNA 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- ...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 Rad50, binds the DNA end, and assembles a DNA cutting channel for the nuclease reactions. The structures reveal how Mre11-Rad50 can detect and process diverse DNA ends and uncover a clamping and gating function for the coiled coils.
External linksMol Cell / PubMed:31492634
MethodsEM (single particle)
Resolution3.5 - 15.6 Å
Structure data

10107

6s6v

EMDB-10107, PDB-6s6v:
Resting state of the E. coli Mre11-Rad50 (SbcCD) head complex bound to ATPgS
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-10114:
The E.coli Mre11-Rad50 complex (SbcCD) bound to ADP and dsDNA with the first coiled-coil interaction site
Method: EM (single particle) / Resolution: 4.3 Å

EMDB-10115:
The E.coli Mre11-Rad50 complex bound to ADP and dsDNA with 200A visible coiled-coils
Method: EM (single particle) / Resolution: 15.6 Å

10116

6s85

EMDB-10116, PDB-6s85:
Cutting state of the E. coli Mre11-Rad50 (SbcCD) head complex bound to ADP and dsDNA.
Method: EM (single particle) / Resolution: 4.2 Å

Chemicals

ChemComp-MN:
Unknown entry

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

Source
  • escherichia coli (E. coli)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN / Nuclease / DNA repair / ABC-type ATPase / DNA double-strand breaks

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