Aurora B regulates MCAK at the mitotic centromere

Dev Cell. 2004 Feb;6(2):253-68. doi: 10.1016/s1534-5807(04)00025-5.

Abstract

Chromosome orientation and alignment within the mitotic spindle requires the Aurora B protein kinase and the mitotic centromere-associated kinesin (MCAK). Here, we report the regulation of MCAK by Aurora B. Aurora B inhibited MCAK's microtubule depolymerizing activity in vitro, and phospho-mimic (S/E) mutants of MCAK inhibited depolymerization in vivo. Expression of either MCAK (S/E) or MCAK (S/A) mutants increased the frequency of syntelic microtubule-kinetochore attachments and mono-oriented chromosomes. MCAK phosphorylation also regulates MCAK localization: the MCAK (S/E) mutant frequently localized to the inner centromere while the (S/A) mutant concentrated at kinetochores. We also detected two different binding sites for MCAK using FRAP analysis of the different MCAK mutants. Moreover, disruption of Aurora B function by expression of a kinase-dead mutant or RNAi prevented centromeric targeting of MCAK. These results link Aurora B activity to MCAK function, with Aurora B regulating MCAK's activity and its localization at the centromere and kinetochore.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antineoplastic Agents, Phytogenic / pharmacology
  • Aurora Kinase B
  • Aurora Kinases
  • Autoradiography
  • CHO Cells
  • Cell Cycle / drug effects
  • Cell Cycle / physiology*
  • Centromere / metabolism*
  • Chromatography, High Pressure Liquid
  • Chromosomes / metabolism
  • Cricetinae
  • Electrophoresis, Gel, Two-Dimensional
  • Fluorescence Recovery After Photobleaching / methods
  • Fluorescent Antibody Technique / methods
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • In Vitro Techniques
  • Kinesin / metabolism*
  • Kinetochores / metabolism
  • Luminescent Proteins / metabolism
  • Mitosis / drug effects
  • Mitosis / physiology*
  • Models, Biological
  • Mutation
  • Nocodazole / pharmacology
  • Paclitaxel / pharmacology
  • Phosphorylation
  • Protein-Serine-Threonine Kinases / metabolism*
  • RNA, Small Interfering / pharmacology
  • Rats
  • Transfection

Substances

  • Antineoplastic Agents, Phytogenic
  • KIF2C protein, human
  • Luminescent Proteins
  • RNA, Small Interfering
  • mitotic centromere-associated kinesin, hamster
  • Green Fluorescent Proteins
  • AURKB protein, human
  • Aurkb protein, rat
  • Aurora Kinase B
  • Aurora Kinases
  • Protein-Serine-Threonine Kinases
  • Kinesin
  • Paclitaxel
  • Nocodazole