Targeting the INCENP IN-box-Aurora B interaction to inhibit CPC activity in vivo.
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.
Wellcome Trust Centre for Cell Biology, Institute of Cell Biology, University of Edinburgh, Michael Swann Building, Kings Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
Florence H
FH
Gohard
Institute for Research in Immunology and Cancer, Department of Medicine, Université de Montréal, Pavillon Marcelle-Coutu, 2950 chemin de Polytechnique, Montréal, Québec H3T 1J4, Canada.
Daniel J
http://orcid.org/0000-0003-2432-6585
DJ
St-Cyr
Wellcome Trust Centre for Cell Biology, Institute of Cell Biology, University of Edinburgh, Michael Swann Building, Kings Buildings, Mayfield Road, Edinburgh EH9 3JR, UK Institute for Research in Immunology and Cancer, Department of Medicine, Université de Montréal, Pavillon Marcelle-Coutu, 2950 chemin de Polytechnique, Montréal, Québec H3T 1J4, Canada.
Mike
http://orcid.org/0000-0002-9713-9994
M
Tyers
Wellcome Trust Centre for Cell Biology, Institute of Cell Biology, University of Edinburgh, Michael Swann Building, Kings Buildings, Mayfield Road, Edinburgh EH9 3JR, UK bill.earnshaw@ed.ac.uk.
William C
WC
Earnshaw
D000595:Amino Acid Sequence; D064107:Aurora Kinase B / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology; D000077022:Survivin
C107489:BIRC5 protein, human; C490157:CDCA8 protein, human; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C083767:INCENP protein, human; D051034:Inhibitor of Apoptosis Proteins; D010455:Peptides; D054852:Small Molecule Libraries; D000077022:Survivin; C576921:AURKB protein, human; D064107:Aurora Kinase B
Aurora B; INCENP; chromosomal passenger complex; cyclic peptide; cytokinesis; mitosis