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

• Published in: Cell reports (Cambridge) 2016-03, Vol.14 (12), p.2975-2987
• Main Authors: Chou, En-Ju, Hung, Liang-Yi, Tang, Chieh-Ju C., Hsu, Wen-Bin, Wu, Hsin-Yi, Liao, Pao-Chi, Tang, Tang K.
• Format: Article
• Language: English
• Subjects: Antigens - genetics; Antigens - metabolism; Aurora Kinase A - genetics; Aurora Kinase A - metabolism; 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
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2016.02.085

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. [Display omitted] •Depletion of CPAP induces multiple mitotic abnormalities•CPAP is phosphorylated by Aurora-A at Ser 467 during mitosis•Phosphorylated CPAP coheres PCM proteins and maintains centrosome integrity•Phosphorylated CPAP has a high affinity for PCM proteins but a low affinity for MTs Chou et al. show that CPAP is essential for proper mitotic progression and maintenance of spindle pole integrity. CPAP is phosphorylated by Aurora-A during mitosis and phosphorylated CPAP coheres PCM proteins and maintains centrosome integrity.