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p14 Arf promotes small ubiquitin-like modifier conjugation of Werners helicase

Yvonne L Woods1, Dimitris P Xirodimas, Alan R Prescott

  • 1CR-UK Cell Transformation Research Group, Department of Surgery and Molecular Oncology, Ninewells Hospital and Medical School, Ninewells Avenue, Dundee DD1 9SY, UK. y.l.woods@dundee.ac.uk

The Journal of Biological Chemistry
|September 10, 2004
PubMed

Insights

The tumor suppressor p14 Arf interacts with Werners helicase (WRN) independently of p53. P14 Arf also promotes WRN SUMOylation, a key step in WRN nuclear redistribution.

Area of Science:

  • Molecular Biology
  • Oncology
  • Cell Biology

Background:

  • p14 Arf and Werners helicase (WRN) are nucleolar tumor suppressors.
  • Their interaction and regulatory mechanisms are not fully understood.
  • p53-independent pathways are crucial in tumor suppression.

Purpose of the Study:

  • To investigate the interaction between p14 Arf and WRN.
  • To elucidate the role of p14 Arf in WRN modification and localization.
  • To determine if SUMOylation is involved in WRN regulation by p14 Arf.

Main Methods:

  • Co-immunoprecipitation assays to study protein interactions.
  • Site-directed mutagenesis to identify binding residues.
  • SUMOylation assays and Western blotting.
  • Confocal microscopy for protein localization studies.

Main Results:

  • A novel, p53-independent interaction between p14 Arf and WRN was demonstrated.
  • Specific residues in p14 Arf (2-14, 82-101) and WRN (central, C-terminus) are critical for binding.
  • p14 Arf synergistically enhances WRN SUMOylation with UBCH9.
  • p14 Arf induces WRN nuclear redistribution, reversed by SUMO-specific protease.

Conclusions:

  • p14 Arf binds multivalently to WRN, similar to its interaction with Mdm2.
  • p14 Arf promotes WRN SUMOylation and nuclear re-localization.
  • SUMOylation is a key mechanism by which p14 Arf regulates WRN.
  • The ability to promote SUMO conjugation is a general function of the p14 Arf tumor suppressor.

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