Microtubules accelerate the kinase activity of Aurora-B by a reduction in dimensionality

PLoS One. 2014 Feb 3;9(2):e86786. doi: 10.1371/journal.pone.0086786. eCollection 2014.

Abstract

Aurora-B is the kinase subunit of the Chromosome Passenger Complex (CPC), a key regulator of mitotic progression that corrects improper kinetochore attachments and establishes the spindle midzone. Recent work has demonstrated that the CPC is a microtubule-associated protein complex and that microtubules are able to activate the CPC by contributing to Aurora-B auto-phosphorylation in trans. Aurora-B activation is thought to occur when the local concentration of Aurora-B is high, as occurs when Aurora-B is enriched at centromeres. It is not clear, however, whether distributed binding to large structures such as microtubules would increase the local concentration of Aurora-B. Here we show that microtubules accelerate the kinase activity of Aurora-B by a "reduction in dimensionality." We find that microtubules increase the kinase activity of Aurora-B toward microtubule-associated substrates while reducing the phosphorylation levels of substrates not associated to microtubules. Using the single molecule assay for microtubule-associated proteins, we show that a minimal CPC construct binds to microtubules and diffuses in a one-dimensional (1D) random walk. The binding of Aurora-B to microtubules is salt-dependent and requires the C-terminal tails of tubulin, indicating that the interaction is electrostatic. We show that the rate of Aurora-B auto-activation is faster with increasing concentrations of microtubules. Finally, we demonstrate that microtubules lose their ability to stimulate Aurora-B when their C-terminal tails are removed by proteolysis. We propose a model in which microtubules act as scaffolds for the enzymatic activity of Aurora-B. The scaffolding activity of microtubules enables rapid Aurora-B activation and efficient phosphorylation of microtubule-associated substrates.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aurora Kinase B / chemistry
  • Aurora Kinase B / genetics
  • Aurora Kinase B / metabolism*
  • Centromere / metabolism*
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation
  • Kinetics
  • Microtubules / chemistry
  • Microtubules / metabolism*
  • Models, Biological
  • Models, Molecular
  • Phosphorylation
  • Protein Binding
  • Protein Structure, Tertiary
  • Substrate Specificity
  • Xenopus Proteins / chemistry
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism*
  • Xenopus laevis / genetics
  • Xenopus laevis / metabolism

Substances

  • Chromosomal Proteins, Non-Histone
  • INCENP protein, Xenopus
  • Xenopus Proteins
  • Aurora Kinase B