Structural mechanism for the recognition of E2F1 by the ubiquitin ligase adaptor Cyclin F

Proc Natl Acad Sci U S A. 2025 Jul;122(26):e2501057122. doi: 10.1073/pnas.2501057122. Epub 2025 Jun 23.

Abstract

Cyclin F, a noncanonical member of the cyclin protein family, plays a critical role in regulating transitions in the cell division cycle. Unlike canonical cyclins, which bind and activate cyclin-dependent kinases (CDKs), Cyclin F functions as a substrate receptor protein within the Skp1-Cullin-F-box E3 ubiquitin ligase complex, enabling the ubiquitylation of target proteins. The structural features that distinguish Cyclin F as a ligase adaptor and the mechanisms underlying its selective substrate recruitment over Cyclin A, which functions in complex with CDK2 at a similar time in the cell cycle, remain largely unexplored. We utilized single-particle cryoelectron microscopy to elucidate the structure of a Cyclin F-Skp1 complex bound to an E2F1 peptide. The structure and biochemical analysis reveal important differences in the substrate-binding site of Cyclin F compared to Cyclin A. Our findings expand on the canonical cyclin-binding motif (Cy or RxL) and highlight the importance of electrostatics at the E2F1 binding interface, which varies between Cyclin F and Cyclin A. These results advance our understanding of E2F1 regulation and may inform strategies for selectively targeting Cyclin F in cancer or neurodegeneration.

Keywords: cell cycle; cryoelectron microscopy; ubiquitin ligase.

MeSH terms

  • Binding Sites
  • Cryoelectron Microscopy
  • Cyclin A / chemistry
  • Cyclin A / metabolism
  • Cyclins* / chemistry
  • Cyclins* / genetics
  • Cyclins* / metabolism
  • E2F1 Transcription Factor* / chemistry
  • E2F1 Transcription Factor* / genetics
  • E2F1 Transcription Factor* / metabolism
  • Humans
  • Models, Molecular
  • Protein Binding
  • S-Phase Kinase-Associated Proteins / chemistry
  • S-Phase Kinase-Associated Proteins / metabolism

Substances

  • E2F1 Transcription Factor
  • Cyclins
  • CCNF protein, human
  • E2F1 protein, human
  • Cyclin A
  • S-Phase Kinase-Associated Proteins
  • SKP1 protein, human