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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

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Phosphorylation of the centrosomal protein, Cep169, by Cdk1 promotes its dissociation from centrosomes in mitosis

Yusuke Mori1, Yoko Inoue1, Yuki Taniyama1

  • 1Department of Chemistry and Biochemistry, School of Advanced Science and Engineering, Waseda University, 3-4-1 Ohkubo, Tokyo 169-8555, Japan.

Biochemical and Biophysical Research Communications
|November 10, 2015
PubMed

Insights

Cep169 protein dissociates from centrosomes during mitosis, a process regulated by Cdk1/Cyclin B. This finding suggests Cep169

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Cep169 is a conserved centrosomal protein crucial for microtubule (MT) regulation.
  • Cep169 collaborates with CDK5RAP2 to stabilize MTs.
  • The exact role of Cep169 in cellular functions, particularly during mitosis, is not fully understood.

Purpose of the Study:

  • To investigate the cell cycle-dependent localization of Cep169.
  • To elucidate the regulatory mechanisms controlling Cep169's centrosomal association.
  • To explore Cep169's potential role in mitotic spindle dynamics.

Main Methods:

  • Immunofluorescence microscopy to track Cep169 localization throughout the cell cycle.
  • Treatment with Cdk1 inhibitor (purvalanol A) to assess its effect on Cep169 localization.
  • Mass spectrometry to identify phosphorylation sites on Cep169.

Main Results:

  • Cep169 localizes to centrosomes during interphase but dissociates at mitosis onset.
  • CDK5RAP2 remains at centrosomes throughout the cell cycle.
  • Cdk1 inhibition prevents Cep169 dissociation from centrosomes during mitosis.
  • Mass spectrometry identified 7 Cdk1 phosphorylation sites on Cep169.

Conclusions:

  • Cep169's dissociation from centrosomes during mitosis is regulated by Cdk1/Cyclin B.
  • Cep169 phosphorylation by Cdk1 controls its mitotic centrosomal localization.
  • Cep169 may play a role in regulating the dynamics of the mitotic spindle MTs.
Keywords:
Cdk1CentrosomeCep169MitosisPhosphorylation

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