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Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
10:27

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

The TRAF6 RING finger domain mediates physical interaction with Ubc13

Jill Wooff1, Landon Pastushok, Michelle Hanna

  • 1Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, SK, Canada S7N 5E5.

FEBS Letters
|May 19, 2004
PubMed

Insights

Tumor necrosis factor receptor associated factor 6 (TRAF6) directly binds to the Ubc13-Uev complex, confirming its role in ubiquitination. This interaction is crucial for TRAF6

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Protein Interactions

Background:

  • Tumor necrosis factor receptor associated factor 6 (TRAF6) is a key signaling protein.
  • TRAF6 functions as a ubiquitin ligase (E3) for Lys-63 polyubiquitination.
  • The interaction between TRAF6 and the Ubc13-Uev (E2) complex was previously uncharacterized.

Purpose of the Study:

  • To investigate the physical interaction between TRAF6 and the Ubc13-Uev E2 complex.
  • To identify the domains involved in the TRAF6-Ubc13 interaction.
  • To map the self-interaction domain of TRAF6.

Main Methods:

  • Yeast two-hybrid assay was employed to detect protein-protein interactions.
  • Site-directed mutagenesis was used to probe the interaction interface.
  • Mapping of self-interaction domains was performed.

Main Results:

  • TRAF6 directly interacts with the Ubc13-Uev complex via binding to Ubc13.
  • Specific mutations in the TRAF6 RING finger domain or Ubc13 surface abolish this interaction.
  • TRAF6 self-interaction was identified and mapped to its N-terminal RING finger motif.

Conclusions:

  • Direct physical interaction between TRAF6 and Ubc13-Uev complex is confirmed.
  • The study provides insights into the molecular interface for Ubc13-TRAF6 RING finger complex formation.
  • Understanding this interaction is vital for elucidating TRAF6-mediated signaling pathways.

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