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The CRL4DCAF1 cullin-RING ubiquitin ligase is activated following a switch in oligomerization state

Weaam I Mohamed1,2, Andreas D Schenk1, Georg Kempf1

  • 1Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.

The EMBO Journal
|October 1, 2021
PubMed

Insights

The CRL4DCAF1 ligase is kept inactive through a novel tetrameric structure that blocks its catalytic site. Neddylation breaks this structure, activating the E3 ubiquitin ligase for cellular processes.

Area of Science:

  • Biochemistry
  • Structural Biology
  • Molecular Cell Biology

Background:

  • CRL4 E3 ubiquitin ligases are crucial for cell survival and growth, with DCAF1 being an essential substrate receptor.
  • Deregulation of CRL4DCAF1 is linked to cancer development.
  • Understanding CRL4DCAF1 structure and mechanism is vital for cellular regulation and disease research.

Purpose of the Study:

  • To investigate the structural basis of CRL4DCAF1 ligase regulation.
  • To elucidate the mechanism controlling CRL4DCAF1 activity.
  • To identify how CRL4DCAF1 is maintained in an inactive state.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) at 8.4 Å resolution.
  • Biochemical assays to study enzyme activity.
  • Structural analysis of CRL4DCAF1 complex.

Main Results:

  • CRL4DCAF1 forms an inactive tetramer where DCAF1's WD40 domain inhibits the RBX1 catalytic subunit of a neighboring protomer.
  • Neddylation disrupts the inhibitory interaction, leading to an active dimeric conformation.
  • A tetramerization-deficient mutant exhibits enhanced ubiquitin ligase activity.

Conclusions:

  • CRL4DCAF1 utilizes a novel autoinhibitory mechanism involving tetramerization to regulate its activity.
  • Neddylation is a key trigger for activating the CRL4DCAF1 ligase.
  • This study reveals how inactive CRL4 ligases are maintained in a substrate-free state.
Keywords:
CRL4/DCAF1E3 ligasesOligomerizationUbiquitinVprBP

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