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TitleMechanism of DNA capture by the MukBEF SMC complex and its inhibition by a viral DNA mimic.
Journal, issue, pagesCell, Year 2025
Publish dateMar 27, 2025
AuthorsFrank Bürmann / Bryony Clifton / Sophie Koekemoer / Oliver J Wilkinson / Dari Kimanius / Mark S Dillingham / Jan Löwe /
PubMed AbstractRing-like structural maintenance of chromosome (SMC) complexes are crucial for genome organization and operate through mechanisms of DNA entrapment and loop extrusion. Here, we explore the DNA ...Ring-like structural maintenance of chromosome (SMC) complexes are crucial for genome organization and operate through mechanisms of DNA entrapment and loop extrusion. Here, we explore the DNA loading process of the bacterial SMC complex MukBEF. Using cryoelectron microscopy (cryo-EM), we demonstrate that ATP binding opens one of MukBEF's three potential DNA entry gates, exposing a DNA capture site that positions DNA at the open neck gate. We discover that the gp5.9 protein of bacteriophage T7 blocks this capture site by DNA mimicry, thereby preventing DNA loading and inactivating MukBEF. We propose a comprehensive and unidirectional loading mechanism in which DNA is first captured at the complex's periphery and then ingested through the DNA entry gate, powered by a single cycle of ATP hydrolysis. These findings illuminate a fundamental aspect of how ubiquitous DNA organizers are primed for genome maintenance and demonstrate how this process can be disrupted by viruses.
External linksCell / PubMed:40168993
MethodsEM (single particle)
Resolution3.7 - 9.1 Å
Structure data

51442

9gm6

EMDB-51442, PDB-9gm6:
MukBEF in a nucleotide-bound state with open neck gate (heads core)
Method: EM (single particle) / Resolution: 3.7 Å

51443

9gm7

EMDB-51443, PDB-9gm7:
MukBEF in a nucleotide-bound state with open neck gate (monomer)
Method: EM (single particle) / Resolution: 4.3 Å

51444

9gm8

EMDB-51444, PDB-9gm8:
MukBEF in a nucleotide-bound state with open neck gate
Method: EM (single particle) / Resolution: 3.9 Å

51445

9gm9

EMDB-51445, PDB-9gm9:
MukBEF in a DNA capture state
Method: EM (single particle) / Resolution: 7.8 Å

51446

9gma

EMDB-51446, PDB-9gma:
MukBEF in a DNA capture state (dimer)
Method: EM (single particle) / Resolution: 9.1 Å

51447

9gmb

EMDB-51447, PDB-9gmb:
MukEF in complex with the phage protein gp5.9
Method: EM (single particle) / Resolution: 4.2 Å

51448

9gmd

EMDB-51448, PDB-9gmd:
MukEF in complex with the phage protein gp5.9 (focus)
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

Source
  • photorhabdus thracensis (bacteria)
  • escherichia coli (E. coli)
  • escherichia phage t7 (virus)
KeywordsDNA BINDING PROTEIN / SMC complex / Chromosome segregation / ABC-type ATPase / DNA mimic

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