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SAS-6 defines a protein family required for centrosome duplication in C. elegans and in human cells

Abstract

The mechanisms that ensure centrosome duplication are poorly understood. In Caenorhabditis elegans, ZYG-1, SAS-4, SAS-5 and SPD-2 are required for centriole formation. However, it is unclear whether these proteins have functional homologues in other organisms. Here, we identify SAS-6 as a component that is required for daughter centriole formation in C. elegans. SAS-6 is a coiled-coil protein that is recruited to centrioles at the onset of the centrosome duplication cycle. Our analysis indicates that SAS-6 and SAS-5 associate and that this interaction, as well as ZYG-1 function, is required for SAS-6 centriolar recruitment. SAS-6 is the founding member of an evolutionarily conserved protein family that contains the novel PISA motif. We investigated the function of the human homologue of SAS-6. GFP–HsSAS-6 localizes to centrosomes and its overexpression results in excess foci-bearing centriolar markers. Furthermore, siRNA-mediated inactivation of HsSAS-6 in U2OS cells abrogates centrosome overduplication following aphidicolin treatment and interferes with the normal centrosome duplication cycle. Therefore, HsSAS-6 is also required for centrosome duplication, indicating that the function of SAS-6-related proteins has been widely conserved during evolution.

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Figure 1: sas-6 is required for daughter centriole formation.
Figure 2: SAS-6 is the founding member of the conserved PISA-containing protein family.
Figure 3: SAS-6 localizes to centrioles.
Figure 4: SAS-6 is recruited to centrioles once per cell cycle.
Figure 5: SAS-6 centriolar targeting.
Figure 6: SAS-6 and SAS-5 physically interact.
Figure 7: HsSAS-6 is required for centrosome duplication in human cells.

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Acknowledgements

We are especially grateful to X. Yan, R. Habedanck and E. Nigg for guidance with cell culture techniques and reagents, as well as M. Cockell for help in generating SAS-6 fusion proteins. We thank J. Azimsadeh, M. Bornens, B. Eddé, M. Glotzer and V. Simanis for reagents. C. Echeverri is acknowledged for communicating results before publication; R. Iggo, V. Simanis and P. Strnad for critical reading of the manuscript; M. Migliaccio, P. Strnad and A. Wilson for help with FACS analysis; C. Bonnard and N. Garin for microscopy support; and D. Moersch and I. Jouravleff for helpful discussions. Some strains were obtained from the Caenorhabditis Genetics Center, which is funded by the National Institute of Health National Center for Research Resources (NCRR). M. D. is recipient of a Roche Research Foundation postdoctoral fellowship. Oncosuisse supports work on centrosome duplication in the Gönczy laboratory (OCS-01495-02-2004).

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Correspondence to Pierre Gönczy.

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Leidel, S., Delattre, M., Cerutti, L. et al. SAS-6 defines a protein family required for centrosome duplication in C. elegans and in human cells. Nat Cell Biol 7, 115–125 (2005). https://doi.org/10.1038/ncb1220

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