Branched microtubule nucleation and dynein transport organize RanGTP asters in Xenopus laevis egg extract

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• dc.contributor.author Scrofani, Jacopo, 1984-
• dc.contributor.author Ruhnow, Felix
• dc.contributor.author Chew, Wei-Xiang
• dc.contributor.author Normanno, Davide
• dc.contributor.author Nédélec, François
• dc.contributor.author Surrey, Thomas
• dc.contributor.author Vernos, Isabelle, 1959-
• dc.date.accessioned 2024-02-06T07:25:16Z
• dc.date.available 2024-02-06T07:25:16Z
• dc.date.issued 2024
• dc.description.abstract Chromosome segregation relies on the correct assembly of a bipolar spindle. Spindle pole self-organization requires dynein-dependent microtubule (MT) transport along other MTs. However, during M-phase RanGTP triggers MT nucleation and branching generating polarized arrays with nonastral organization in which MT minus ends are linked to the sides of other MTs. This raises the question of how branched-MT nucleation and dynein-mediated transport cooperate to organize the spindle poles. Here, we used RanGTP-dependent MT aster formation in Xenopus laevis (X. laevis) egg extract to study the interplay between these two seemingly conflicting organizing principles. Using temporally controlled perturbations of MT nucleation and dynein activity, we found that branched MTs are not static but instead dynamically redistribute over time as poles self-organize. Our experimental data together with computer simulations suggest a model where dynein together with dynactin and NuMA directly pulls and move branched MT minus ends toward other MT minus ends.
• dc.description.sponsorship We acknowledge Dr. Andreas Merdes (CBI, Toulouse; France) for the anti-NuMA serum. We thank the CRG microscopy facility for training, microscope maintenance and technical support. Work in Vernos lab was supported by grants from the Spanish Ministry of Economy (MINECO) I+D grants BFU2015-68726-P and by European Regional Development Fund (ERDF), the Ministry of Science, Innovation and Universities grant PGC2018-096976-B-I00 funded by Ministerio de Ciencia e Innovación (MCIN)/Agencia. Estatal de Investigació (AEI)/10.13039/501100011033/ and by ERDF A way of doing Europe, grant Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR)-SGR 478 from the Generalitat de Catalunya and intramural funds from the Centre for Genomic Regulation. Work in the Surrey lab was supported by the Spanish Ministry of Economy (MINECO) grant PID2019-108415GB-I00/ AEI/10.13039/501100011033. This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme under grant agreement No 951430, also supporting F.N. W.X.C. is supported by a Human Frontier Science Program fellowship (HFSP LT000682/2020-C). We acknowledge support of the Spanish Ministry of Science and Innovation through the Centro de Excelencia Severo Ochoa (CEX2020-001049-S, MCIN/AEI/10.13039/501100011033), and the Generalitat de Catalunya through the CERCA programme. We are grateful to the CRG Core Technologies Programme for their support and assistance in this work.
• dc.format.mimetype application/pdf
• dc.identifier.citation Scrofani J, Ruhnow F, Chew WX, Normanno D, Nedelec F, Surrey T, Vernos I. Branched microtubule nucleation and dynein transport organize RanGTP asters in Xenopus laevis egg extract. Mol Biol Cell. 2024 Jan 1;35(1):ar12. DOI: 10.1091/mbc.E23-10-0407
• dc.identifier.doi http://dx.doi.org/10.1091/mbc.E23-10-0407
• dc.identifier.issn 1059-1524
• dc.identifier.uri http://hdl.handle.net/10230/58963
• dc.language.iso eng
• dc.publisher American Society for Cell Biology
• dc.relation.ispartof Mol Biol Cell. 2024 Jan 1;35(1):ar12
• dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/951430
• dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/BFU2015-68726-P
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PGC2018-096976-B-I00
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PID2019-108415GB-I00
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/CEX2020-001049-S
• dc.rights © 2024 Scrofani et al. This article is distributed by The American Society for Cell Biology under license from the author(s). It is available to the public under an Attribution 4.0 International Creative Commons CC-BY 4.0 License (https://creativecommons.org/licenses/by/4.0/).
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.other Cèl·lules--Divisió
• dc.title Branched microtubule nucleation and dynein transport organize RanGTP asters in Xenopus laevis egg extract
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion