Clathrin recruits phosphorylated TACC3 to spindle poles for bipolar spindle assembly and chromosome alignment

J Cell Sci. 2010 Nov 1;123(Pt 21):3645-51. doi: 10.1242/jcs.075911. Epub 2010 Oct 5.

Abstract

Transforming acidic coiled-coil-containing protein 3 (TACC3) has been implicated in mitotic spindle assembly, although the mechanisms involved are largely unknown. Here we identify that clathrin heavy chain (CHC) binds specifically to phosphorylated TACC3 and recruits it to spindle poles for proper spindle assembly and chromosome alignment. Phosphorylation of Xenopus TACC3 at serine 620 (S620) and S626, but not S33, is required for its binding with CHC. Knockdown of CHC by RNA interference (RNAi) abolishes the targeting of TACC3 to spindle poles and results in abnormal spindle assembly and chromosome misalignment, similar to the defects caused by TACC3 knockdown. Furthermore, the binding of CHC with phosphorylated TACC3 is inhibited by importin β and this inhibition is reversed by the presence of the GTP-binding nuclear protein Ran in the GTP-bound state. Together, these results indicate that the recruitment of phosphorylated TACC3 to spindle poles by CHC ensures proper spindle assembly and chromosome alignment, and is regulated by Ran.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aurora Kinases
  • Chromosomes / metabolism
  • Chromosomes / ultrastructure
  • Clathrin Heavy Chains / metabolism*
  • HeLa Cells
  • Humans
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Phosphorylation
  • Protein Binding / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Protein Transport / genetics
  • RNA, Small Interfering / genetics
  • Spindle Apparatus / metabolism*
  • Xenopus laevis
  • beta Karyopherins / metabolism*
  • ran GTP-Binding Protein / metabolism*

Substances

  • Microtubule-Associated Proteins
  • RNA, Small Interfering
  • TACC3 protein, human
  • beta Karyopherins
  • Clathrin Heavy Chains
  • Aurora Kinases
  • Protein Serine-Threonine Kinases
  • ran GTP-Binding Protein