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Cited by 35DNA Repair MechanismsGenomics and Chromatin DynamicsMicrotubule and mitosis dynamics

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Cryo-EM structure of the Smc5/6 holo-complex

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Hallett, Stephen T, Campbell Harry, Isabella, Schellenberger, Pascale, Zhou, Lihong, Cronin, Nora B, Baxter, Jonathan, Etheridge, Thomas J, Murray, Johanne M, Oliver, Antony W·2022·Nucleic Acids Research
DOI 10.1093/nar/gkac692PMC PMC9458440

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Abstract

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.

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Metadata

Authors
Hallett, S. T., Campbell Harry, I., Schellenberger, P., Zhou, L., Cronin, N. B., Baxter, J., Etheridge, T. J., Murray, J. M., & Oliver, A. W.
Affiliations
  • University of Sussex (GB)
  • Genome Damage and Stability Centre, School of Life Sciences, University of Sussex , Falmer , UK
  • The Francis Crick Institute (GB)
Corresponding author
  • Etheridge, Thomas J
  • Murray, Johanne M
  • Oliver, Antony W
ORCID
Venue
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Published
2022-08-22
Volume
50
Issue
16
Pages
9505-9520
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
DOI
10.1093/nar/gkac692
PMC
PMC9458440
OpenAlex
W4292607806
Cited by
35
References
2
Citations by year
2026: 10 · 2025: 5 · 2024: 9 · 2023: 8 · 2022: 3
FWCI
2.04
Type
journal article
Language
en
Open access
Licence
Locations
  • Nucleic Acids Research (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Sussex Research Online (University of Sussex) (submittedVersion, cc-by)
  • Figshare (submittedVersion, cc-by)
  • The Francis Crick Institute (cc-by)
Funders
  • Wellcome Trust (095605/Z/11/A)
  • University of Sussex
  • Wellcome (110047/Z/15/Z)
  • Cancer Research UK (C302/A24386)
  • MRC (MR/PO18955/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
Keywords
Smc5/6 complex, SMC complex, SMC5, SMC4, SMC1A, mitotic recombination, meiotic recombination, replication stress, viral replication restriction, Saccharomyces cerevisiae, protein complex structure
MeSH terms
Chromosomal Proteins, Non-Histone, Nuclear Proteins, Saccharomyces cerevisiae, Cell Cycle Proteins, Cryoelectron Microscopy, Saccharomyces cerevisiae Proteins

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