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MUL1-RING recruits the substrate, p53-TAD as a complex with UBE2D2-UB conjugate

Min-Sung Lee1,2, Sang-Ok Lee1,3, Joonhyeok Choi4

  • 1Disease Target Structure Research Center, Division of Biomedical Research, KRIBB, Daejeon, South Korea.

The FEBS Journal
|January 20, 2022
PubMed

Insights

The MUL1 RING domain recruits UBE2D2 and p53 simultaneously, enabling ubiquitylation. This involves UBE2D2 conformational changes and enhanced p53 binding, revealing a multivalent interaction mechanism.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • The MUL1 RING domain (RINGMUL1) mediates ubiquitylation of the p53-transactivation domain (TADp53).
  • Understanding the recruitment mechanism of UBE2D2 and TADp53 by RINGMUL1 is crucial for elucidating ubiquitylation pathways.

Purpose of the Study:

  • To determine the structural basis for the simultaneous recruitment of UBE2D2 and TADp53 by RINGMUL1.
  • To investigate the conformational changes in UBE2D2 upon binding to RINGMUL1 and its effect on TADp53 interaction.

Main Methods:

  • Complex structure determination of RINGMUL1:UBE2D2.
  • Interaction studies using UBE2D2-UB thioester mimetics and TADp53.
  • Analysis of UBE2D2 mutations (S22R/C85S, N77A) and their impact on conformation and hydrolysis.

Main Results:

  • RINGMUL1 binding induces a closed conformation of UBE2D2RS-UBROE and accelerates its hydrolysis, an effect modulated by the N77A mutation.
  • UBE2D2RAS-UBROE adopts a closed conformation even without RINGMUL1.
  • TADp53 binding affinity increases significantly for the RINGMUL1:UBE2D2 and RINGMUL1:UBE2D2RAS-UBROE complexes.

Conclusions:

  • The simultaneous recognition of TADp53 by RINGMUL1 in complex with UBE2D2~UB is driven by multivalent binding events.
  • This multivalent interaction mechanism underlies the ability of RINGMUL1 to ubiquitylate the intrinsically disordered TADp53.
Keywords:
MUL1 RING domainNMRUBE2D2p53 transactivation domainubiquitin

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