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First published online 14 August 2007
doi: 10.1242/jcs.010322

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Ninein is released from the centrosome and moves bi-directionally along microtubules

David K. Moss^1,*, Gemma Bellett¹, Jane M. Carter¹, Mirjana Liovic^2,3, Jennifer Keynton¹, Alan R. Prescott⁴, E. Birgitte Lane² and Mette M. Mogensen^1,

¹ School of Biological Sciences, University of East Anglia, Norwich, NR4 7TJ, UK
² Division of Cell and Developmental Biology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK
³ Medical Centre for Molecular Biology, Faculty of Medicine University of Ljubljana, Vrazov trg 2, SI-1000 Ljubljana, Slovenia
⁴ Division of Cell Biology and Immunology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK

Author for correspondence (e-mail: m.mogensen{at}uea.ac.uk)

Accepted 26 June 2007

Cell-to-cell contact and polarisation of epithelial cells involve a major reorganisation of the microtubules and centrosomal components. The radial microtubule organisation is lost and an apico-basal array develops that is no longer anchored at the centrosome. This involves not only the relocation of microtubules but also of centrosomal anchoring proteins to apical non-centrosomal sites. The relocation of microtubule minus-end-anchoring proteins such as ninein to the apical sites is likely to be essential for the assembly and stabilisation of the apico-basal arrays in polarised epithelial cells. In this study, we establish that ninein is highly dynamic and that, in epithelial cells, it is present not only at the centrosome but also in the cytoplasm as distinct speckles. Live-cell imaging reveals that GFP-ninein speckles are released from the centrosome and move in a microtubule-dependent manner within the cytoplasm and thus establishes that epithelial cells possess the mechanical means for relocation of ninein to non-centrosomal anchoring sites. We also provide evidence for the deployment of ninein speckles to apical anchoring sites during epithelial differentiation in both an in situ tissue and an in vitro culture system. In addition, the findings suggest that the non-centrosomal microtubule anchoring sites associate with adherens junctions in polarised epithelial cells.

Key words: Centrosome, Ninein, Polarised epithelial cells, Cochlea, Apico-basal microtubule arrays

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