Phosphorylation of CPAP by Aurora-A Maintains Spindle Pole Integrity during Mitosis

Cell Rep. 2016 Mar 29;14(12):2975-87. doi: 10.1016/j.celrep.2016.02.085. Epub 2016 Mar 17.

Abstract

CPAP is required for centriole elongation during S/G2 phase, but the role of CPAP in mitosis is incompletely understood. Here, we show that CPAP maintains spindle pole integrity through its phosphorylation by Aurora-A during mitosis. Depletion of CPAP induced a prolonged delay in mitosis, pericentriolar material (PCM) dispersion, and multiple mitotic abnormalities. Further studies demonstrated that CPAP directly interacts with and is phosphorylated by Aurora-A at serine 467 during mitosis. Interestingly, the dispersal of the PCM was effectively rescued by ectopic expression of wild-type CPAP or a phospho-mimic CPAP-S467D mutant, but not a non-phosphorylated CPAP-S467A mutant. Finally, we found that CPAP-S467D has a low affinity for microtubule binding but a high affinity for PCM proteins. Together, our results support a model wherein CPAP is required for proper mitotic progression, and phosphorylation of CPAP by Aurora-A is essential for maintaining spindle pole integrity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens / genetics
  • Antigens / metabolism
  • Aurora Kinase A / genetics
  • Aurora Kinase A / metabolism*
  • Cell Cycle Proteins
  • Centrosome / physiology
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Microscopy, Confocal
  • Microtubule-Associated Proteins / antagonists & inhibitors
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Mitosis*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Phosphorylation
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Spindle Poles / physiology*
  • Time-Lapse Imaging
  • Tubulin / genetics
  • Tubulin / metabolism

Substances

  • Antigens
  • CDK5RAP2 protein, human
  • CENPJ protein, human
  • Cell Cycle Proteins
  • Intracellular Signaling Peptides and Proteins
  • Microtubule-Associated Proteins
  • Nerve Tissue Proteins
  • RNA, Small Interfering
  • Tubulin
  • pericentrin
  • Aurora Kinase A