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MBC in Press, published online ahead of print July 11, 2003
Mol. Biol. Cell 10.1091/mbc.E02-11-0773

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Submitted on November 28, 2002
Revised on June 6, 2003
Accepted on June 6, 2003

Part of Ran is associated with AKAP450 at the centrosome. Involvement in microtubule organizing activity

Guy Keryer, Barbara Di Fiore1, Claude Celati, Karl Ferdinand Lechtreck2, Mette Mogensen3, Annie Delouvée, Patrizia Lavia1, Michel Bornens, and Anne-Marie Tassin*

1 Institute of Molecular Biology and Pathology-CNR c/o Università "La Sapienza", Rome (Italy)
2 Department of Botany, University of Cologne (Germany)
3 School of Biological Sciences, University of East Anglia, Norwich, (UK)

* Corresponding author. E-mail address: atassin{at}curie.fr.

The small Ran GTPase, a key regulator of nucleocytoplasmic transport is also involved in microtubule assembly and nuclear membrane formation. Here we show by immunofluorescence, immuno-EM and biochemical analysis that a fraction of Ran is tightly associated with the centrosome throughout the cell cycle. Ran interaction with the centrosome is mediated by the centrosomal matrix A Kinase Anchoring Protein (AKAP450). Accordingly, when AKAP450 is delocalized from the centrosome, Ran is also delocalized and, as a consequence, microtubule regrowth or anchoring is altered, despite the persisting association of {gamma}-tubulin with the centrosome. Moreover Ran is recruited to Xenopus sperm centrosome during its activation for microtubule nucleation. We also demonstrate that centrosomal proteins such as centrin and pericentrin but not {gamma}-tubulin, AKAP450, or ninein, undertake a nucleocytoplasmic exchange as they concentrate in the nucleus upon export inhibition by leptomycin B. Altogether these results suggest a challenging possibility namely that centrosome activity could depend upon nucleocytoplasmic exchange of centrosomal proteins and local Ran-dependent concentration at the centrosome.




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