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In Vitro Analysis of E3 Ubiquitin Ligase Function
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In Vitro Analysis of E3 Ubiquitin Ligase Function

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CYLD Regulates Centriolar Satellites Proteostasis by Counteracting the E3 Ligase MIB1

Tiphaine Douanne1, Gwennan André-Grégoire2, An Thys1

  • 1CRCINA, Team SOAP, INSERM, CNRS, Université de Nantes, Université d'Angers, IRS-UN blg, Room 405, 8 quai Moncousu, 44007 Nantes, France.

Cell Reports
|May 9, 2019
PubMed

Insights

The tumor suppressor CYLD deubiquitinates proteins, aiding primary cilia formation. CYLD prevents the degradation of centriolar satellites by targeting the E3 ligase MIB1, thus regulating ciliogenesis.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The tumor suppressor CYLD is a deubiquitinating enzyme crucial for signal transduction.
  • CYLD plays a role in primary cilia formation, acting as a cellular sensing antenna.
  • The precise mechanisms by which CYLD regulates ciliogenesis remain incompletely understood.

Purpose of the Study:

  • To identify CYLD binding partners involved in ciliogenesis.
  • To elucidate the role of CYLD in regulating centriolar satellites and proteostasis.
  • To understand how CYLD influences the ubiquitination status of key ciliogenesis factors.

Main Methods:

  • Unbiased proteomic screening to identify CYLD interacting proteins.
  • Analysis of centriolar satellite components, including PCM1.
  • Investigation of the E3 ligase Mindbomb 1 (MIB1) and its role in PCM1 degradation.
  • Assessment of CYLD knockdown effects on PCM1 stability and centriolar satellites.

Main Results:

  • Proteomic screening identified centriolar satellite components as CYLD binding partners.
  • CYLD knockdown led to PCM1 degradation and centriolar satellite disassembly.
  • CYLD was found to deubiquitinate and inhibit MIB1-mediated proteasomal degradation of PCM1.
  • These findings establish a link between CYLD, centriolar satellite proteostasis, and ciliogenesis.

Conclusions:

  • CYLD is essential for maintaining the stability of centriolar satellites by regulating PCM1 proteostasis.
  • CYLD functions by deubiquitinating and inhibiting the E3 ligase MIB1, preventing PCM1 degradation.
  • This study provides novel insights into the regulation of ciliogenesis through reversible ubiquitination.
Keywords:
CYLDMIB1PCM1centriolar satellitesciliogenesisproteostasisubiquitin

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