A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes
The spindle position checkpoint (SPOC) is a spindle pole body (SPB, equivalent of mammalian centrosome) associated surveillance mechanism that halts mitotic exit upon spindle mis-orientation. Here, we monitored the interaction between SPB proteins and the SPOC component Bfa1 by FRET microscopy. We s...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2016-05-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/14029 |
| _version_ | 1811201038013693952 |
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| author | Yuliya Gryaznova Ayse Koca Caydasi Gabriele Malengo Victor Sourjik Gislene Pereira |
| author_facet | Yuliya Gryaznova Ayse Koca Caydasi Gabriele Malengo Victor Sourjik Gislene Pereira |
| author_sort | Yuliya Gryaznova |
| collection | DOAJ |
| description | The spindle position checkpoint (SPOC) is a spindle pole body (SPB, equivalent of mammalian centrosome) associated surveillance mechanism that halts mitotic exit upon spindle mis-orientation. Here, we monitored the interaction between SPB proteins and the SPOC component Bfa1 by FRET microscopy. We show that Bfa1 binds to the scaffold-protein Nud1 and the γ-tubulin receptor Spc72. Spindle misalignment specifically disrupts Bfa1-Spc72 interaction by a mechanism that requires the 14-3-3-family protein Bmh1 and the MARK/PAR-kinase Kin4. Dissociation of Bfa1 from Spc72 prevents the inhibitory phosphorylation of Bfa1 by the polo-like kinase Cdc5. We propose Spc72 as a regulatory hub that coordinates the activity of Kin4 and Cdc5 towards Bfa1. In addition, analysis of spc72∆ cells shows that a mitotic-exit-promoting dominant signal, which is triggered upon elongation of the spindle into the bud, overrides the SPOC. Our data reinforce the importance of daughter-cell-associated factors and centrosome-based regulations in mitotic exit and SPOC control. |
| first_indexed | 2024-04-12T02:15:08Z |
| format | Article |
| id | doaj.art-f1a059a336494e2890f1cd400d1c5dbb |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:15:08Z |
| publishDate | 2016-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-f1a059a336494e2890f1cd400d1c5dbb2022-12-22T03:52:17ZengeLife Sciences Publications LtdeLife2050-084X2016-05-01510.7554/eLife.14029A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomesYuliya Gryaznova0Ayse Koca Caydasi1Gabriele Malengo2Victor Sourjik3Gislene Pereira4https://orcid.org/0000-0002-6519-4737DKFZ-ZMBH Alliance, German Cancer Research Centre, Heidelberg, GermanyDKFZ-ZMBH Alliance, German Cancer Research Centre, Heidelberg, Germany; Centre for Organismal Studies, University of Heidelberg, Heidelberg, GermanyDKFZ-ZMBH Alliance, Centre for Molecular Biology, Heidelberg, GermanyDKFZ-ZMBH Alliance, Centre for Molecular Biology, Heidelberg, GermanyDKFZ-ZMBH Alliance, German Cancer Research Centre, Heidelberg, Germany; Centre for Organismal Studies, University of Heidelberg, Heidelberg, GermanyThe spindle position checkpoint (SPOC) is a spindle pole body (SPB, equivalent of mammalian centrosome) associated surveillance mechanism that halts mitotic exit upon spindle mis-orientation. Here, we monitored the interaction between SPB proteins and the SPOC component Bfa1 by FRET microscopy. We show that Bfa1 binds to the scaffold-protein Nud1 and the γ-tubulin receptor Spc72. Spindle misalignment specifically disrupts Bfa1-Spc72 interaction by a mechanism that requires the 14-3-3-family protein Bmh1 and the MARK/PAR-kinase Kin4. Dissociation of Bfa1 from Spc72 prevents the inhibitory phosphorylation of Bfa1 by the polo-like kinase Cdc5. We propose Spc72 as a regulatory hub that coordinates the activity of Kin4 and Cdc5 towards Bfa1. In addition, analysis of spc72∆ cells shows that a mitotic-exit-promoting dominant signal, which is triggered upon elongation of the spindle into the bud, overrides the SPOC. Our data reinforce the importance of daughter-cell-associated factors and centrosome-based regulations in mitotic exit and SPOC control.https://elifesciences.org/articles/14029checkpointcentrosome/spindle pole bodymitosis |
| spellingShingle | Yuliya Gryaznova Ayse Koca Caydasi Gabriele Malengo Victor Sourjik Gislene Pereira A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes eLife checkpoint centrosome/spindle pole body mitosis |
| title | A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes |
| title_full | A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes |
| title_fullStr | A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes |
| title_full_unstemmed | A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes |
| title_short | A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes |
| title_sort | fret based study reveals site specific regulation of spindle position checkpoint proteins at yeast centrosomes |
| topic | checkpoint centrosome/spindle pole body mitosis |
| url | https://elifesciences.org/articles/14029 |
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