Abstract
The centrosome is the major microtubule-organizing centre of many cells, best known for its role in mitotic spindle organization. How the proteins of the centrosome are accurately assembled to carry out its many functions remains poorly understood. The non-membrane-bound nature of the centrosome dictates that protein–protein interactions drive its assembly and functions. To investigate this massive macromolecular organelle, we generated a ‘domain-level' centrosome interactome using direct protein–protein interaction data from a focused yeast two-hybrid screen. We then used biochemistry, cell biology and the model organism Drosophila to provide insight into the protein organization and kinase regulatory machinery required for centrosome assembly. Finally, we identified a novel role for Plk4, the master regulator of centriole duplication. We show that Plk4 phosphorylates Cep135 to properly position the essential centriole component Asterless. This interaction landscape affords a critical framework for research of normal and aberrant centrosomes.
Highlights
The centrosome is the major microtubule-organizing centre of many cells, best known for its role in mitotic spindle organization
These interactions are likely modified by regulated changes in protein-binding affinity in a cell-type and cell-cycle-dependent manner, which can in turn modulate centrosome behaviour and function
This study focused on identifying protein–protein interactions (PPIs) among a core set of conserved centrosome proteins, including proteins of the centriole, the pericentriolar material (PCM) and regulatory kinases
Summary
The centrosome is the major microtubule-organizing centre of many cells, best known for its role in mitotic spindle organization. The non-membranebound nature of the centrosome dictates that protein–protein interactions drive its assembly and functions. We show that Plk[4] phosphorylates Cep[135] to properly position the essential centriole component Asterless This interaction landscape affords a critical framework for research of normal and aberrant centrosomes. A much smaller set of proteins was identified as essential for centrosome duplication or activity through genetic analysis in model systems[6] and high-throughput RNA interference screens in culture[7,8,9]. There is a poor understanding of how centrosome proteins are assembled to form this structure and carry out its functions at the level of direct protein–protein interactions (PPIs). New methods for identifying PPIs are critical for moving forward Another poorly understood aspect of centrosome regulation is the precise role of regulatory kinases. Identifying novel substrates for these kinases, and other centrosome kinases such as Nek[2], Aurora A and LK6, will uncover how they exert influence on centrosome assembly and function
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