CaMKIIgamma-mediated inactivation of the Kin I kinesin MCAK is essential for bipolar spindle formation

EMBO J. 2005 Mar 23;24(6):1256-66. doi: 10.1038/sj.emboj.7600601. Epub 2005 Mar 3.

Abstract

MCAK, a member of the kinesin-13 family, is a microtubule (MT) depolymerase that is necessary to ensure proper kinetochore MT attachment during spindle formation. Regulation of MCAK activity and localization is controlled in part by Aurora B kinase at the centromere. Here we analyzed human cells depleted of the ubiquitous Ca(2+)/calmodulin-dependent protein kinase IIgamma isoform (CaMKIIgamma) by RNA interference and found that CaMKIIgamma was necessary to suppress MCAK depolymerase activity in vivo. A functional overlap with TOGp, a MT regulator known to counteract MCAK, was suggested by similar CaMKIIgamma- and TOGp-depletion phenotypes, namely disorganized multipolar spindles. A replicating vector system, which permits inducible overexpression in cells that simultaneously synthesize interfering short hairpin RNAs, was used to dissect the functional interplay between CaMKIIgamma, TOGp, and MCAK. Our results revealed two distinct but functionally overlapping mechanisms for negative regulation of the cytosolic/centrosomal pool of MCAK. These two mechanisms, involving CaMKIIgamma and TOGp, respectively, are both essential for spindle bipolarity in a normal physiological context, but not in MCAK-depleted cells.

Publication types

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

MeSH terms

  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases / genetics
  • Calcium-Calmodulin-Dependent Protein Kinases / physiology*
  • Cell Division / genetics
  • Cell Division / physiology
  • Humans
  • Kinesin / metabolism*
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / physiology
  • Nucleic Acid Conformation
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Spindle Apparatus / metabolism*

Substances

  • CKAP5 protein, human
  • KIF2C protein, human
  • Microtubule-Associated Proteins
  • RNA, Small Interfering
  • CAMK2G protein, human
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Kinesin