Pericentrin-mediated SAS-6 recruitment promotes centriole assembly.

Daisuke Ito,José B Pereira-Leal,Miguel Godinho Ferreira,Swadhin Chandra Jana,Zita Carvalho-Santos,Jaroslaw Surkont,Sihem Zitouni,Paulo Duarte,Mónica Bettencourt-Dias

doi:10.7554/elife.41418

Abstract

The centrosome is composed of two centrioles surrounded by a microtubule-nucleating pericentriolar material (PCM). Although centrioles are known to regulate PCM assembly, it is less known whether and how the PCM contributes to centriole assembly. Here we investigate the interaction between centriole components and the PCM by taking advantage of fission yeast, which has a centriole-free, PCM-containing centrosome, the SPB. Surprisingly, we observed that several ectopically-expressed animal centriole components such as SAS-6 are recruited to the SPB. We revealed that a conserved PCM component, Pcp1/pericentrin, interacts with and recruits SAS-6. This interaction is conserved and important for centriole assembly, particularly its elongation. We further explored how yeasts kept this interaction even after centriole loss and showed that the conserved calmodulin-binding region of Pcp1/pericentrin is critical for SAS-6 interaction. Our work suggests that the PCM not only recruits and concentrates microtubule-nucleators, but also the centriole assembly machinery, promoting biogenesis close by.

Highlights

The centrosome is the major microtubule organizing center found in animals, being composed of two microtubule-based cylinders, the centrioles, which are surrounded by an electron-dense proteinaceous pericentriolar material (PCM) that nucleates microtubules
We first investigated the conservation of centrosome components, searching for orthologues of the known animal proteins comprising: components of centrioles that are required for centriole biogenesis (SAS-6, STIL/Ana2/SAS-5, CPAP/SAS-4, CEP135/Bld10 and CEP295/Ana1), linkers of the centriole to the PCM, which are bound to the centriole and are required for PCM recruitment (CEP152 and CEP192), and the PCM itself, which is involved in g-tubulin recruitment and anchoring
We examined the relationship between the PCM and centriole components, taking advantage of a heterologous system, the fission yeast, which does not have centrioles, and has lost the coding sequences for its components

Summary

Introduction

The centrosome is the major microtubule organizing center found in animals, being composed of two microtubule-based cylinders, the centrioles, which are surrounded by an electron-dense proteinaceous pericentriolar material (PCM) that nucleates microtubules. The first structure of the centriole to be assembled is the cartwheel, a ninefold symmetrical structure, composed of SAS-6, CEP135/Bld, STIL/Ana2/SAS-5, amongst others (Kitagawa et al, 2011; Lin et al, 2013; Nakazawa et al, 2007; Tang et al, 2011; van Breugel et al, 2011). This is followed by centriole elongation through the deposition of centriolar microtubules which is dependent on components such as CPAP/SAS-4 (Kohlmaier et al, 2009; Schmidt et al, 2009; Tang et al, 2009). CDK5RAP2/CNN and pericentrin are recruited, which recruit and activate g-tubulin and create an environment favorable for concentrating

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Conclusion