Abstract
At the onset of mitosis, the centrosome undergoes maturation, which is characterized by a drastic expansion of the pericentriolar material (PCM) and a robust increase in microtubule-organizing activity. CEP215 is one of the major PCM components which accumulates at the centrosome during mitosis. The depletion phenotypes indicate that CEP215 is essential for centrosome maturation and bipolar spindle formation. Here, we performed a series of knockdown-rescue experiments to link the protein-protein interaction properties of CEP215 to its biological functions. The results showed that CEP215 and pericentrin, another major PCM component, is interdependent for their accumulation at the spindle poles during mitosis. As a result, The CEP215-pericentrin interaction is required for centrosome maturation and subsequent bipolar spindle formation during mitosis. On the other hand, CEP215 interaction with γ-tubulin is dispensable for centrosome maturation. Our results provide an insight how PCM components are assembled to form a spindle pole during mitosis.
Highlights
The centrosome in most animal cells functions as a major microtubule organizing center and controls cellular morphology, migration and subcellular transport
CEP215 Interaction with Pericentrin for its Localization and Spindle Pole Formation In this work, we examined importance of CEP215-pericentrin interaction in mitotic cells
The centrosomal pericentrin levels were still not recovered with FLAG-CEP215DC-PACT (Fig. 1C and E)
Summary
The centrosome in most animal cells functions as a major microtubule organizing center and controls cellular morphology, migration and subcellular transport. The amount of PCM and microtubule organizing activity of the centrosome fluctuate in parallel during the cell cycle. At the onset of mitosis, the centrosome matures with a robust increase in centrosomal ctubulin. The failure of this process causes defects in bipolar spindle formation and chromosome congression. Selected PCM components, including pericentrin [1,2], CEP215/CDK5RAP2 [3,4], NEDD1/GCP-WD [5,6] and CEP192 [7,8] are known to be critical for c-tubulin recruitment to the centrosome. The means by which these PCM components accumulate and organize to form a mature centrosome is not fully understood
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