Cryo-EM structures of apo-APC/C and APC/CCDH1:EMI1 complexes provide insights into APC/C regulation

Nat Commun. 2024 Nov 21;15(1):10074. doi: 10.1038/s41467-024-54398-5.

Abstract

APC/C is a multi-subunit complex that functions as a master regulator of cell division. It controls progression through the cell cycle by timely marking mitotic cyclins and other cell cycle regulatory proteins for degradation. The APC/C itself is regulated by the sequential action of its coactivator subunits CDC20 and CDH1, post-translational modifications, and its inhibitory binding partners EMI1 and the mitotic checkpoint complex. In this study, we took advantage of developments in cryo-electron microscopy to determine the structures of human APC/CCDH1:EMI1 and apo-APC/C at 2.9 Å and 3.2 Å resolution, respectively, providing insights into the regulation of APC/C activity. The high-resolution maps allow the unambiguous assignment of an α-helix to the N-terminus of CDH1 (CDH1α1) in the APC/CCDH1:EMI1 ternary complex. We also identify a zinc-binding module in APC2 that confers structural stability to the complex, and we confirm the presence of zinc ions experimentally. Finally, due to the higher resolution and well defined density of these maps, we are able to build, aided by AlphaFold predictions, several intrinsically disordered regions in different APC/C subunits that likely play a role in proper APC/C assembly and regulation of its activity.

MeSH terms

  • Anaphase-Promoting Complex-Cyclosome* / chemistry
  • Anaphase-Promoting Complex-Cyclosome* / metabolism
  • Anaphase-Promoting Complex-Cyclosome* / ultrastructure
  • Antigens, CD
  • Cadherins / chemistry
  • Cadherins / metabolism
  • Cadherins / ultrastructure
  • Cdc20 Proteins / chemistry
  • Cdc20 Proteins / metabolism
  • Cdc20 Proteins / ultrastructure
  • Cdh1 Proteins / chemistry
  • Cdh1 Proteins / genetics
  • Cdh1 Proteins / metabolism
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / metabolism
  • Cell Cycle Proteins / ultrastructure
  • Cryoelectron Microscopy*
  • Humans
  • Models, Molecular
  • Protein Binding
  • Zinc / chemistry
  • Zinc / metabolism

Substances

  • Anaphase-Promoting Complex-Cyclosome
  • CDH1 protein, human
  • Zinc
  • Cell Cycle Proteins
  • Cdh1 Proteins
  • Cadherins
  • Cdc20 Proteins
  • Antigens, CD