Requirement of aurora-A kinase in astral microtubule polymerization and spindle microtubule flux

Cell Cycle. 2008 Apr 15;7(8):1104-11. doi: 10.4161/cc.7.8.5738. Epub 2008 Feb 11.

Abstract

Mitotic Aurora-A kinase was found to be required for formation of bipolar spindle, ensuring accurate chromosome segregation in mitosis. Recently, Aurora-A was shown to promote Ran-GTP-induced spindle formation and astral microtubule development. Here, by selective immunodepletion, we showed that Aurora-A was required for centrosome- but not Ran-GTP-induced astral microtubule formation in Xenopus egg extracts. Aurora-A enhanced microtubule polymerization in both centrosome- and Ran-GTP-induced aster assemblies: shortening the timing of aster assembly and increasing the aster size. Indeed, adding of Aurora-A protein alone induced microtubule clustering, which was abrogated by Aurora kinase inhibitory small molecule ZM447439. In addition, we showed that Aurora-A was indispensable for Ran-GTP-induced bipolar spindle formation. Inhibition of Aurora-A activity by adding of kinase inactive dominant mutant led to spindle collapse and formation of monopolar spindle whereas minus-end motor protein dynein/dynactin inhibitor p50/dynamitin rescued the bipolar structure. Lastly, we revealed that Aurora-A was necessary for microtubule poleward flux and this requirement depended on kinase activity. Thus, we showed that Aurora-A promoted microtubule polymerization and maintained microtubule flux in ensuring proper bipolar spindle assembly.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aurora Kinases
  • Benzamides
  • Biological Transport / physiology
  • Chromosome Segregation / physiology*
  • Immunoblotting
  • Microtubules / metabolism*
  • Polymers
  • Protein Serine-Threonine Kinases / metabolism*
  • Quinazolines
  • Spindle Apparatus / metabolism*
  • Xenopus

Substances

  • 4-(4-(N-benzoylamino)anilino)-6-methoxy-7-(3-(1-morpholino)propoxy)quinazoline
  • Benzamides
  • Polymers
  • Quinazolines
  • Aurora Kinases
  • Protein Serine-Threonine Kinases