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P21-activated kinase 4 (PAK4) is required for metaphase spindle positioning and anchoring

Oncogene volume 32, pages 910–919 (2013)Cite this article

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Abstract

The oncogenic kinase PAK4 was recently found to be involved in the regulation of the G1 phase and the G2/M transition of the cell cycle. We have also identified that PAK4 regulates Ran GTPase activity during mitosis. Here, we show that after entering mitosis, PAK4-depleted cells maintain a prolonged metaphase-like state. In these cells, chromosome congression to the metaphase plate occurs with normal kinetics but is followed by an extended period during which membrane blebbing and spindle rotation are observed. These bipolar PAK4-depleted metaphase-like spindles have a defective astral microtubule (MT) network and are not centered in the cell but are in close contact with the cell cortex. As the metaphase-like state persists, centrosome fragmentation occurs, chromosomes scatter from the metaphase plate and move toward the spindle poles with an active spindle assembly checkpoint, a phenotype that is reminiscent of cohesion fatigue. PAK4 also regulates the acto-myosin cytoskeleton and we report that PAK4 depletion results in the induction of cortical membrane blebbing during prometaphase arrest. However, we show that membrane blebs, which are strongly enriched in phospho-cofilin, are not responsible for the poor anchoring of the spindle. As PAK4 depletion interferes with the localization of components of the dynein/dynactin complexes at the kinetochores and on the astral MTs, we propose that loss of PAK4 could induce a change in the activities of motor proteins.

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Abbreviations

PAK4:

p21-activated kinase 4

MT:

microtubules

KT:

kinetochore

SAC:

spindle assembly checkpoint.

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Acknowledgements

We thank Drs Lorca, Castro, Merdes for the gift of reagents. We are indebted to Dr A Desai, Dr K Vaughan and Dr A Musacchio for the kind gifts of reagents. We want to especially thank Dr Dan Fisher (IGMM, UMR5535 CNRS, Montpellier, France) for critical reading and editing of the manuscript. We thank the Montpellier RIO imaging facility. GB is supported by a grant from ‘Fondation de la Recherche Médicale’. This work was supported by a grant MEGAPAK to NM from the ANR (Agence Nationale pour la Recherche) GENOPAT.

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Author notes

  1. G Bompard

    Present address: 5Current address: IGH, CNRS UPR1142, 141, rue de la Cardonille, 34396 Montpellier, France,

Authors and Affiliations

  1. Universités Montpellier 2 et 1, Montpellier, France

    G Bompard, G Rabeharivelo, J Cau, A Abrieu, C Delsert & N Morin

  2. CRBM, CNRS, UMR 5237, Montpellier, France

    G Bompard, G Rabeharivelo, A Abrieu, C Delsert & N Morin

  3. MRI, IGH, CNRS UPR1142, Montpellier, France

    J Cau

  4. IFREMER, LGP, la, Tremblade, France

    C Delsert

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  1. G Bompard
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Correspondence to N Morin.

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Bompard, G., Rabeharivelo, G., Cau, J. et al. P21-activated kinase 4 (PAK4) is required for metaphase spindle positioning and anchoring. Oncogene 32, 910–919 (2013). https://doi.org/10.1038/onc.2012.98

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