Phosphorylation of maskin by Aurora-A is regulated by RanGTP and importin beta

J Biol Chem. 2006 Dec 15;281(50):38293-301. doi: 10.1074/jbc.M607203200. Epub 2006 Oct 22.

Abstract

Mitotic spindle assembly in Xenopus egg extracts is regulated at least in part by importin beta and its regulator, the small GTPase, Ran. RanGTP stabilizes microtubules near the chromosomes during spindle assembly by selectively releasing spindle assembly factors from inhibition by importin alpha/beta in the vicinity of the chromosomes. Several spindle assembly factors are regulated in this manner. We identified maskin, the Xenopus member of the transforming acidic coiled coil family of proteins, as a potential candidate in a two-step affinity chromatography approach designed to uncover additional downstream targets of importin alpha/beta in mitosis. Here, we show that although maskin lacks a canonical nuclear localization sequence, it binds importin beta in a RanGTP-regulated manner. We further show that importin beta inhibits the regulatory phosphorylation of maskin by Aurora-A. This suggests a novel mechanism by which importin beta regulates the activity of a spindle assembly factor.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aurora Kinases
  • Base Sequence
  • Chromatography, Affinity
  • DNA Primers
  • Phosphorylation
  • Protein Serine-Threonine Kinases / metabolism*
  • Transcription Factors / metabolism*
  • Xenopus Proteins / metabolism*
  • beta Karyopherins / metabolism*
  • ran GTP-Binding Protein / metabolism*

Substances

  • DNA Primers
  • TACC3 protein, Xenopus
  • Transcription Factors
  • Xenopus Proteins
  • beta Karyopherins
  • Aurora Kinases
  • Protein Serine-Threonine Kinases
  • ran GTP-Binding Protein