Targeting the INCENP IN-box-Aurora B interaction to inhibit CPC activity in vivo

Open Biol. 2014 Nov;4(11):140163. doi: 10.1098/rsob.140163.

Abstract

The chromosome passenger complex (CPC) is an essential regulator of mitosis and cytokinesis. The CPC consists of Aurora B kinase, inner centromere protein (INCENP), and the targeting subunits survivin and borealin/Dasra B. INCENP is a scaffolding subunit for the CPC and activates Aurora B via its conserved IN-box domain. We show that overexpression of soluble IN-box in HeLa cells affects endogenous CPC localization and produces a significant increase in multinucleated and micronucleated cells consistent with CPC loss of function. The dominant-negative effect of soluble IN-box expression depends on residues corresponding to hINCENP W845 and/or F881, suggesting that these are essential for Aurora B binding in vivo. We then screened a targeted library of small (five to nine residues long) circular peptide (CP) IN-box fragments generated using split intein circular ligation of proteins and peptides (SICLOPPS) methodology. We identified a number of CPs that caused modest but reproducible increases in rates of multinucleated and micronucleated cells. Our results provide proof of concept that inhibition of the Aurora B-IN-box interaction is a viable strategy for interfering with CPC function in vivo.

Keywords: Aurora B; INCENP; chromosomal passenger complex; cyclic peptide; cytokinesis; mitosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aurora Kinase B / chemistry
  • Aurora Kinase B / genetics
  • Aurora Kinase B / metabolism*
  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • HeLa Cells
  • Humans
  • Inhibitor of Apoptosis Proteins / metabolism
  • Molecular Sequence Data
  • Peptides / isolation & purification
  • Peptides / pharmacology
  • Protein Binding / drug effects
  • Protein Structure, Tertiary
  • Protein Transport
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology
  • Survivin

Substances

  • BIRC5 protein, human
  • CDCA8 protein, human
  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • INCENP protein, human
  • Inhibitor of Apoptosis Proteins
  • Peptides
  • Small Molecule Libraries
  • Survivin
  • AURKB protein, human
  • Aurora Kinase B