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dc.contributor.authorGouveia, Susana Montenegro-
dc.contributor.authorZitouni, Sihem-
dc.contributor.authorKong, Dong-
dc.contributor.authorDuarte, Paulo-
dc.contributor.authorGomes, Beatriz Ferreira-
dc.contributor.authorSousa, Ana Laura-
dc.contributor.authorTranfield, Erin M.-
dc.contributor.authorHyman, Anthony-
dc.contributor.authorLoncarek, Jadranka-
dc.contributor.authorBettencourt-Dias, Monica-
dc.descriptionThe 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.pt_PT
dc.descriptionThe deposited article version contains attached the supplementary materials within the pdf.pt_PT
dc.description.abstractThe 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.pt_PT
dc.description.sponsorshipWe are thankful to Anna Akhmanova, Raquel Oliveira and Jeffrey B.Woodruff for reading and discussing the manuscript. We are also thankful to Catarina Nabais for the GFP control construct and Vladimir Joukov for the Xenopus Cep192 antibody. S.M.G was funded by an EMBO Long term fellowship ALTF 1088-2009, a Marie curie Intra-European fellowship (#253373) and a FCT postdoctoral fellowship. The collaboration with J.L. laboratory in the USA was supported by a The Company of Biologists travel grant. S.Z is funded by ERC grant ERC-COG-683258. Research in JL lab was supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. M.B-D. Laboratory is supported by an ERC grant ERC-COG-683258 and FCT Investigator to MBD.pt_PT
dc.publisherCompany of Biologistspt_PT
dc.relationALTF 1088-2009pt_PT
dc.subjectin vitro reconstitutionpt_PT
dc.subjectmicrotubule nucleationpt_PT
dc.subjectde novopt_PT
dc.subjectsupramolecular assembliespt_PT
dc.titlePLK4 is a microtubule-associated protein that self assembles promoting de novo MTOC formationpt_PT
degois.publication.titleJournal of Cell Sciencept_PT
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