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TitleCryo-EM structure of the Smc5/6 holo-complex.
Journal, issue, pagesNucleic Acids Res, Vol. 50, Issue 16, Page 9505-9520, Year 2022
Publish dateSep 9, 2022
AuthorsStephen T Hallett / Isabella Campbell Harry / Pascale Schellenberger / Lihong Zhou / Nora B Cronin / Jonathan Baxter / Thomas J Etheridge / Johanne M Murray / Antony W Oliver /
PubMed AbstractThe Smc5/6 complex plays an essential role in the resolution of recombination intermediates formed during mitosis or meiosis, or as a result of the cellular response to replication stress. It also ...The Smc5/6 complex plays an essential role in the resolution of recombination intermediates formed during mitosis or meiosis, or as a result of the cellular response to replication stress. It also functions as a restriction factor preventing viral replication. Here, we report the cryogenic EM (cryo-EM) structure of the six-subunit budding yeast Smc5/6 holo-complex, reconstituted from recombinant proteins expressed in insect cells - providing both an architectural overview of the entire complex and an understanding of how the Nse1/3/4 subcomplex binds to the hetero-dimeric SMC protein core. In addition, we demonstrate that a region within the head domain of Smc5, equivalent to the 'W-loop' of Smc4 or 'F-loop' of Smc1, mediates an important interaction with Nse1. Notably, mutations that alter the surface-charge profile of the region of Nse1 which accepts the Smc5-loop, lead to a slow-growth phenotype and a global reduction in the chromatin-associated fraction of the Smc5/6 complex, as judged by single molecule localisation microscopy experiments in live yeast. Moreover, when taken together, our data indicates functional equivalence between the structurally unrelated KITE and HAWK accessory subunits associated with SMC complexes.
External linksNucleic Acids Res / PubMed:35993814 / PubMed Central
MethodsEM (single particle)
Resolution6.5 - 8.53 Å
Structure data

EMDB-13893: Cryo-EM structure of the Smc5/6 holo-complex; map for head-end of complex.
Method: EM (single particle) / Resolution: 6.5 Å

EMDB-13894: CryoEM structure of the Smc5/6 holocomplex; map for hinge and arm region.
Method: EM (single particle) / Resolution: 8.53 Å

EMDB-13895, PDB-7qcd:
CryoEM structure of the Smc5/6-holocomplex (composite structure)
Method: EM (single particle) / Resolution: 8.0 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • Saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
KeywordsRECOMBINATION / Structural Maintenance of Chromosomes / SMC / holo-complex / stalled replication fork

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