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Structure paper

TitleCryo-EM structure of the Mre11-Rad50-Nbs1 complex reveals the molecular mechanism of scaffolding functions.
Journal, issue, pagesMol Cell, Vol. 83, Issue 2, Page 167-185.e9, Year 2023
Publish dateJan 19, 2023
AuthorsMatthias Rotheneder / Kristina Stakyte / Erik van de Logt / Joseph D Bartho / Katja Lammens / Yilan Fan / Aaron Alt / Brigitte Kessler / Christophe Jung / Wynand P Roos / Barbara Steigenberger / Karl-Peter Hopfner /
PubMed AbstractThe DNA double-strand break repair complex Mre11-Rad50-Nbs1 (MRN) detects and nucleolytically processes DNA ends, activates the ATM kinase, and tethers DNA at break sites. How MRN can act both as ...The DNA double-strand break repair complex Mre11-Rad50-Nbs1 (MRN) detects and nucleolytically processes DNA ends, activates the ATM kinase, and tethers DNA at break sites. How MRN can act both as nuclease and scaffold protein is not well understood. The cryo-EM structure of MRN from Chaetomium thermophilum reveals a 2:2:1 complex with a single Nbs1 wrapping around the autoinhibited Mre11 nuclease dimer. MRN has two DNA-binding modes, one ATP-dependent mode for loading onto DNA ends and one ATP-independent mode through Mre11's C terminus, suggesting how it may interact with DSBs and intact DNA. MRNs two 60-nm-long coiled-coil domains form a linear rod structure, the apex of which is assembled by the two joined zinc-hook motifs. Apices from two MRN complexes can further dimerize, forming 120-nm spanning MRN-MRN structures. Our results illustrate the architecture of MRN and suggest how it mechanistically integrates catalytic and tethering functions.
External linksMol Cell / PubMed:36577401
MethodsEM (single particle) / X-ray diffraction
Resolution2.51 - 7.57 Å
Structure data

EMDB-14877: Chaetomium thermophilum MRN complex (Nbs1)
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-14878: CtMre11-Rad50-Nbs1 complex, long coiled-coils map
Method: EM (single particle) / Resolution: 4.75 Å

EMDB-14879: Mre11-Rad50-Nbs1 complex (Chaetomium thermophilum) lower coiled-coils
Method: EM (single particle) / Resolution: 7.57 Å

EMDB-14880: Eukaryotic Mre11-Rad50-Nbs1 complex catalytic head
Method: EM (single particle) / Resolution: 4.0 Å

14881

7zr1

EMDB-14881, PDB-7zr1:
Chaetomium thermophilum Mre11-Rad50-Nbs1 complex bound to ATPyS (composite structure)
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-14882: Chaetomium thermophilum Rad50 zinc-hook tetramer
Method: EM (single particle) / Resolution: 4.9 Å

15948

8bah

EMDB-15948, PDB-8bah:
Human Mre11-Nbs1 complex
Method: EM (single particle) / Resolution: 4.13 Å

PDB-7zqy:
Chaetomium thermophilum Rad50 Zn hook
Method: X-RAY DIFFRACTION / Resolution: 2.51 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-HOH:
WATER / Water

ChemComp-MN:
Unknown entry

ChemComp-MG:
Unknown entry

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

Source
  • Chaetomium thermophilum (fungus)
  • thermochaetoides thermophila (fungus)
  • homo sapiens (human)
KeywordsHYDROLASE / DNA repair / ATPase / coiled coil / Zn-hook / complex / coiled-coils

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