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

Oncogene. 2013 Feb 14;32(7):910-9. doi: 10.1038/onc.2012.98. Epub 2012 Mar 26.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle / genetics
  • Centrosome / metabolism
  • Chromosome Segregation / drug effects
  • Chromosome Segregation / genetics
  • Chromosome Segregation / physiology
  • Cytoplasm / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Kinetochores / metabolism
  • Metaphase* / drug effects
  • Metaphase* / genetics
  • Metaphase* / physiology
  • Microtubules / drug effects
  • Microtubules / metabolism
  • RNA, Small Interfering / pharmacology
  • Spindle Apparatus / drug effects
  • Spindle Apparatus / genetics
  • Spindle Apparatus / metabolism*
  • Spindle Apparatus / physiology
  • Transfection
  • p21-Activated Kinases / antagonists & inhibitors
  • p21-Activated Kinases / genetics
  • p21-Activated Kinases / metabolism
  • p21-Activated Kinases / physiology*

Substances

  • RNA, Small Interfering
  • Green Fluorescent Proteins
  • PAK4 protein, human
  • p21-Activated Kinases