CAMSAPs and nucleation-promoting factors control microtubule release from γ-TuRC
© 2024. The Author(s)..
γ-Tubulin ring complex (γ-TuRC) is the major microtubule-nucleating factor. After nucleation, microtubules can be released from γ-TuRC and stabilized by other proteins, such as CAMSAPs, but the biochemical cross-talk between minus-end regulation pathways is poorly understood. Here we reconstituted this process in vitro using purified components. We found that all CAMSAPs could bind to the minus ends of γ-TuRC-attached microtubules. CAMSAP2 and CAMSAP3, which decorate and stabilize growing minus ends but not the minus-end tracking protein CAMSAP1, induced microtubule release from γ-TuRC. CDK5RAP2, a γ-TuRC-interactor, and CLASP2, a regulator of microtubule growth, strongly stimulated γ-TuRC-dependent microtubule nucleation, but only CDK5RAP2 suppressed CAMSAP binding to γ-TuRC-anchored minus ends and their release. CDK5RAP2 also improved selectivity of γ-tubulin-containing complexes for 13- rather than 14-protofilament microtubules in microtubule-capping assays. Knockout and overexpression experiments in cells showed that CDK5RAP2 inhibits the formation of CAMSAP2-bound microtubules detached from the microtubule-organizing centre. We conclude that CAMSAPs can release newly nucleated microtubules from γ-TuRC, whereas nucleation-promoting factors can differentially regulate this process.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:26 |
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| Enthalten in: |
Nature cell biology - 26(2024), 3 vom: 15. März, Seite 404-420 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Rai, Dipti [VerfasserIn] |
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| Anmerkungen: |
Date Completed 18.03.2024 Date Revised 18.03.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1038/s41556-024-01366-2 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM369141652 |
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| 520 | |a γ-Tubulin ring complex (γ-TuRC) is the major microtubule-nucleating factor. After nucleation, microtubules can be released from γ-TuRC and stabilized by other proteins, such as CAMSAPs, but the biochemical cross-talk between minus-end regulation pathways is poorly understood. Here we reconstituted this process in vitro using purified components. We found that all CAMSAPs could bind to the minus ends of γ-TuRC-attached microtubules. CAMSAP2 and CAMSAP3, which decorate and stabilize growing minus ends but not the minus-end tracking protein CAMSAP1, induced microtubule release from γ-TuRC. CDK5RAP2, a γ-TuRC-interactor, and CLASP2, a regulator of microtubule growth, strongly stimulated γ-TuRC-dependent microtubule nucleation, but only CDK5RAP2 suppressed CAMSAP binding to γ-TuRC-anchored minus ends and their release. CDK5RAP2 also improved selectivity of γ-tubulin-containing complexes for 13- rather than 14-protofilament microtubules in microtubule-capping assays. Knockout and overexpression experiments in cells showed that CDK5RAP2 inhibits the formation of CAMSAP2-bound microtubules detached from the microtubule-organizing centre. We conclude that CAMSAPs can release newly nucleated microtubules from γ-TuRC, whereas nucleation-promoting factors can differentially regulate this process | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 7 | |a Microtubule-Associated Proteins |2 NLM | |
| 700 | 1 | |a Song, Yinlong |e verfasserin |4 aut | |
| 700 | 1 | |a Hua, Shasha |e verfasserin |4 aut | |
| 700 | 1 | |a Stecker, Kelly |e verfasserin |4 aut | |
| 700 | 1 | |a Monster, Jooske L |e verfasserin |4 aut | |
| 700 | 1 | |a Yin, Victor |e verfasserin |4 aut | |
| 700 | 1 | |a Stucchi, Riccardo |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Yixin |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Yaqian |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Fangrui |e verfasserin |4 aut | |
| 700 | 1 | |a Katrukha, Eugene A |e verfasserin |4 aut | |
| 700 | 1 | |a Altelaar, Maarten |e verfasserin |4 aut | |
| 700 | 1 | |a Heck, Albert J R |e verfasserin |4 aut | |
| 700 | 1 | |a Wieczorek, Michal |e verfasserin |4 aut | |
| 700 | 1 | |a Jiang, Kai |e verfasserin |4 aut | |
| 700 | 1 | |a Akhmanova, Anna |e verfasserin |4 aut | |
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