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Interaction of the growth and tumour suppressor NORE1A with microtubules is not required for its growth-suppressive

Anna Moshnikova1, Sergey Kuznetsov, Andrei V Khokhlatchev

  • 1Department of Pathology, University of Virginia, PO Box 800904, Charlottesville, VA 22908-0904, USA. ak9k@virginia.edu

BMC Research Notes
|August 20, 2008
PubMed
Abstract

Insights

Microtubule-associated protein 1B (MAP1B) mediates NORE1A binding to microtubules, but this interaction is not essential for NORE1A

Area of Science:

  • Cell biology
  • Cancer research
  • Molecular mechanisms of cell signaling

Background:

  • NORE1A is a Ras effector and tumor suppressor inactivated in various cancers.
  • NORE1A localizes to microtubules and centrosomes in transformed cells.
  • The mechanism of NORE1A-cytoskeletal association and its role in tumor suppression are unknown.

Purpose of the Study:

  • To elucidate the mechanism of NORE1A association with microtubules and centrosomes.
  • To determine if microtubule or centrosomal localization of NORE1A is required for its tumor-suppressive function.

Main Methods:

  • In vitro microtubule binding assays.
  • Mass spectrometry to identify NORE1A interacting partners.
  • RNA interference (RNAi) to suppress protein expression.
  • Immunodepletion assays.

Main Results:

  • Purified NORE1A did not bind microtubules in vitro, indicating mediation by other proteins.
  • Mass spectrometry identified MAP1B and C19ORF5 as NORE1A interactors.
  • MAP1B, but not C19ORF5, is required for NORE1A-microtubule association.
  • NORE1A's centrosomal localization and tumor-suppressive activity were independent of MAP1B and C19ORF5.

Conclusions:

  • Microtubule-associated protein 1B (MAP1B) mediates NORE1A interaction with microtubules.
  • NORE1A centrosomal localization involves a mechanism distinct from microtubule binding.
  • Microtubule localization of NORE1A is not essential for its growth-suppressive function.

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