Gouveia, Susana MontenegroZitouni, SihemKong, DongDuarte, PauloGomes, Beatriz FerreiraSousa, Ana LauraTranfield, Erin M.Hyman, AnthonyLoncarek, JadrankaBettencourt-Dias, Monica2018-09-252019-09-012018-09-20http://jcs.biologists.org/content/early/2018/09/17/jcs.219501.longhttp://hdl.handle.net/10400.7/896The deposited article version is the Epub Ahead of Print version of the article (the "Accepted Manuscript"), posted online 20th September 2018, provided by Company of Biologists. It has peer-review.The deposited article version contains attached the supplementary materials within the pdf.The centrosome is an important microtubule-organizing centre (MTOC) in animal cells. It consists of two barrel-shaped structures, the centrioles, surrounded by the pericentriolar material (PCM), which nucleates microtubules. Centrosomes can form close to an existing structure (canonical duplication) or de novo How centrosomes form de novo is not known. The master driver of centrosome biogenesis, PLK4, is critical to recruit several centriole components. Here, we investigate the beginning of centrosome biogenesis, taking advantage of Xenopus egg extracts, where PLK4 can induce de novo MTOC formation (Eckerdt et al., 2011; Zitouni et al., 2016). Surprisingly, we observe that in vitro, PLK4 can self-assemble into condensates that recruit α/β-tubulin. In Xenopus extracts, PLK4 assemblies additionally recruit PLK4's substrate, STIL, and the microtubule nucleator, γ-tubulin, forming acentriolar MTOCs de novo The assembly of these robust microtubule asters is independent of dynein, similarly to centrosomes. We suggest a new mechanism of action for PLK4, where it forms a self-organizing catalytic scaffold that recruits centriole components, PCM factors and α/β-tubulin, leading to MTOC formation.engPLK4MTOCsin vitro reconstitutionmicrotubule nucleationPCMcentrosomesde novosupramolecular assembliesPLK4 is a microtubule-associated protein that self assembles promoting de novo MTOC formationjournal article10.1242/jcs.219501