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CPAP interacts with 14-3-3 in a cell cycle-dependent manner.
Chiung-Ya Chen1, Monilola A Olayioye, Geoffrey J Lindeman
1Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan, ROC.
Biochemical and Biophysical Research Communications
|March 7, 2006
Summary
Centrosomal Protein 4.1-Associated Protein (CPAP) interacts with 14-3-3 proteins, a binding dependent on phosphorylation at serine 1109 and regulated by the cell cycle.
Area of Science:
- Cell Biology
- Molecular Biology
- Protein Interactions
Background:
- Centrosomal Protein 4.1-Associated Protein (CPAP) has a known microtubule-destabilizing motif.
- The regulation of microtubule dynamics at the centrosome is crucial for cell division.
Purpose of the Study:
- To identify conserved sequence motifs in CPAP.
- To investigate the interaction between CPAP and 14-3-3 proteins.
- To determine the regulation of CPAP-14-3-3 interaction during the cell cycle.
Main Methods:
- Sequence motif analysis
- Yeast two-hybrid assays
- Co-immunoprecipitation experiments
- Site-directed mutagenesis
- Alkaline phosphatase treatment
Main Results:
- Two classical 14-3-3 binding sites were identified in CPAP.
- CPAP directly interacts with 14-3-3 proteins in a phosphorylation-dependent manner, specifically at serine 1109.
- This interaction is significantly reduced during mitosis, indicating cell cycle regulation.
Conclusions:
- Phosphorylation of CPAP on serine 1109 is essential for 14-3-3 binding.
- The CPAP-14-3-3 interaction is dynamically regulated throughout the cell cycle.
- This interaction likely plays a role in regulating microtubule dynamics at the centrosome.