The centrosome is the main microtubule organization centre of animal cells. It is composed of a centriole pair surrounded by pericentriolar material (PCM). Traditionally described as amorphous, the architecture of the PCM is not known, although its intricate mode of assembly alludes to the presence of a functional, hierarchical structure. Here we used subdiffraction imaging to reveal organizational features of the PCM. Interphase PCM components adopt a concentric toroidal distribution of discrete diameter around centrioles. Positional mapping of multiple non-overlapping epitopes revealed that pericentrin (PCNT) is an elongated molecule extending away from the centriole. We find that PCM components occupy separable spatial domains within mitotic PCM that are maintained in the absence of microtubule nucleation complexes and further implicate PCNT and CDK5RAP2 in the organization and assembly of PCM. Globally, this work highlights the role of higher-order PCM organization in the regulation of centrosome assembly and function.
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We would like to thank members of the Pelletier laboratory for stimulating discussions during the course of this work, especially C. Yeh and J. Gonçalves for their critical reading of the manuscript, C. Holley for esiRNA production, and M. Bashkurov for help with super-resolution imaging. Furthermore, we are grateful to D. Drechsel (MPI-CBG, Dresden, Germany), S. Doxsey (University of Massachusetts Medical School, Worcester, USA), M. Gomez-Ferreria (CRG, Barcelona, Spain), J. Lüders (IRB, Barcelona, Spain), K. Rhee (Seoul National University, Korea), J. Salisbury (Mayo Clinic, Minnesota, USA) and L-H. Tsai (MIT, Cambridge, USA) for providing key reagents and Applied Precision/GE Healthcare for excellent technical support with the OMX microscope. This work was financially supported by the Canadian Cancer Society (019562), the Natural Sciences and Engineering Research Council of Canada (RGPIN-355644-2008) and a grant-in-aid from the Krembil Foundation. L.P. holds a Canada Research Chair (Tier 2) in Centrosome Biogenesis and Function. S.L. is a Vanier Canada Graduate Scholar (CIHR).
The authors declare no competing financial interests.
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Lawo, S., Hasegan, M., Gupta, G. et al. Subdiffraction imaging of centrosomes reveals higher-order organizational features of pericentriolar material. Nat Cell Biol 14, 1148–1158 (2012). https://doi.org/10.1038/ncb2591
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