Cargando…

The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin

Global regulation of spindle-associated proteins is crucial in oocytes due to the absence of centrosomes and their very large cytoplasmic volume, but little is known about how this is achieved beyond involvement of the Ran-importin pathway. We previously uncovered a novel regulatory mechanism in Dro...

Descripción completa

Detalles Bibliográficos
Autores principales: Repton, Charlotte, Cullen, C. Fiona, Costa, Mariana F. A., Spanos, Christos, Rappsilber, Juri, Ohkura, Hiroyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9203013/
https://www.ncbi.nlm.nih.gov/pubmed/35666772
http://dx.doi.org/10.1371/journal.pgen.1009995
_version_ 1784728641474658304
author Repton, Charlotte
Cullen, C. Fiona
Costa, Mariana F. A.
Spanos, Christos
Rappsilber, Juri
Ohkura, Hiroyuki
author_facet Repton, Charlotte
Cullen, C. Fiona
Costa, Mariana F. A.
Spanos, Christos
Rappsilber, Juri
Ohkura, Hiroyuki
author_sort Repton, Charlotte
collection PubMed
description Global regulation of spindle-associated proteins is crucial in oocytes due to the absence of centrosomes and their very large cytoplasmic volume, but little is known about how this is achieved beyond involvement of the Ran-importin pathway. We previously uncovered a novel regulatory mechanism in Drosophila oocytes, in which the phospho-docking protein 14-3-3 suppresses microtubule binding of Kinesin-14/Ncd away from chromosomes. Here we report systematic identification of microtubule-associated proteins regulated by 14-3-3 from Drosophila oocytes. Proteins from ovary extract were co-sedimented with microtubules in the presence or absence of a 14-3-3 inhibitor. Through quantitative mass-spectrometry, we identified proteins or complexes whose ability to bind microtubules is suppressed by 14-3-3, including the chromosomal passenger complex (CPC), the centralspindlin complex and Kinesin-14/Ncd. We showed that 14-3-3 binds to the disordered region of Borealin, and this binding is regulated differentially by two phosphorylations on Borealin. Mutations at these two phospho-sites compromised normal Borealin localisation and centromere bi-orientation in oocytes, showing that phospho-regulation of 14-3-3 binding is important for Borealin localisation and function.
format Online
Article
Text
id pubmed-9203013
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-92030132022-06-17 The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin Repton, Charlotte Cullen, C. Fiona Costa, Mariana F. A. Spanos, Christos Rappsilber, Juri Ohkura, Hiroyuki PLoS Genet Research Article Global regulation of spindle-associated proteins is crucial in oocytes due to the absence of centrosomes and their very large cytoplasmic volume, but little is known about how this is achieved beyond involvement of the Ran-importin pathway. We previously uncovered a novel regulatory mechanism in Drosophila oocytes, in which the phospho-docking protein 14-3-3 suppresses microtubule binding of Kinesin-14/Ncd away from chromosomes. Here we report systematic identification of microtubule-associated proteins regulated by 14-3-3 from Drosophila oocytes. Proteins from ovary extract were co-sedimented with microtubules in the presence or absence of a 14-3-3 inhibitor. Through quantitative mass-spectrometry, we identified proteins or complexes whose ability to bind microtubules is suppressed by 14-3-3, including the chromosomal passenger complex (CPC), the centralspindlin complex and Kinesin-14/Ncd. We showed that 14-3-3 binds to the disordered region of Borealin, and this binding is regulated differentially by two phosphorylations on Borealin. Mutations at these two phospho-sites compromised normal Borealin localisation and centromere bi-orientation in oocytes, showing that phospho-regulation of 14-3-3 binding is important for Borealin localisation and function. Public Library of Science 2022-06-06 /pmc/articles/PMC9203013/ /pubmed/35666772 http://dx.doi.org/10.1371/journal.pgen.1009995 Text en © 2022 Repton et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Repton, Charlotte
Cullen, C. Fiona
Costa, Mariana F. A.
Spanos, Christos
Rappsilber, Juri
Ohkura, Hiroyuki
The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin
title The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin
title_full The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin
title_fullStr The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin
title_full_unstemmed The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin
title_short The phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger Borealin
title_sort phospho-docking protein 14-3-3 regulates microtubule-associated proteins in oocytes including the chromosomal passenger borealin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9203013/
https://www.ncbi.nlm.nih.gov/pubmed/35666772
http://dx.doi.org/10.1371/journal.pgen.1009995
work_keys_str_mv AT reptoncharlotte thephosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT cullencfiona thephosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT costamarianafa thephosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT spanoschristos thephosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT rappsilberjuri thephosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT ohkurahiroyuki thephosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT reptoncharlotte phosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT cullencfiona phosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT costamarianafa phosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT spanoschristos phosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT rappsilberjuri phosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin
AT ohkurahiroyuki phosphodockingprotein1433regulatesmicrotubuleassociatedproteinsinoocytesincludingthechromosomalpassengerborealin