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Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain

Keijun Kakihara1,2, Kengo Asamizu1, Kei Moritsugu3

  • 1School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.

Communications Biology
|November 9, 2021
PubMed

Insights

Ubiquitin-specific protease 8 (USP8) autoinhibition is mediated by a WW-like domain. This region

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Ubiquitin-specific protease 8 (USP8) regulates membrane trafficking.
  • USP8 activity is inhibited by 14-3-3 proteins.
  • USP8 mutations are linked to Cushing's disease.

Purpose of the Study:

  • To elucidate the molecular mechanism of USP8 autoinhibition.
  • To understand how 14-3-3 proteins regulate USP8 activity.
  • To investigate the structural basis of USP8 regulation.

Main Methods:

  • Pull-down assays
  • Single-molecule Förster Resonance Energy Transfer (smFRET)
  • In silico modeling

Main Results:

  • Amino acids 645-684 of USP8 identified as an autoinhibitory region.
  • This region forms a WW-like domain that obstructs the catalytic cleft.
  • 14-3-3 binding enhances the interaction between the WW-like and USP domains, inhibiting activity.

Conclusions:

  • The WW-like domain of USP8 mediates autoinhibition.
  • 14-3-3 proteins further inhibit USP8 by stabilizing this autoinhibitory interaction.
  • Dysregulation of this autoinhibition may contribute to Cushing's disease.

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