A network of interacting ciliary tip proteins with opposing activities imparts slow and processive microtubule growth
Abstract Cilia are essential motile or sensory organelles found on many eukaryotic cells. Their formation and function rely on axonemal microtubules, which exhibit very slow dynamics, however the underlying biochemical mechanisms are largely unexplored. Here, we reconstitutedin vitrothe individual and collective activities of the ciliary tip module proteins, CEP104, CSPP1, TOGARAM1, ARMC9 and CCDC66, which interact with each other and with microtubules, and, when mutated, cause ciliopathies such as Joubert syndrome. CEP104, a protein containing a tubulin-binding TOG domain, is an inhibitor of microtubule growth and shortening that interacts with EBs on the microtubule surface and with a luminal microtubule-pausing factor CSPP1. Another TOG-domain protein, TOGARAM1, overcomes growth inhibition imposed by CEP104 and CSPP1. CCDC66 and ARMC9 do not affect microtubule dynamics directly but act as scaffolds for their partners. Cryo-electron tomography showed that together, ciliary tip module members form plus-end-specific cork-like structures which reduce protofilament flaring. The combined effect of these proteins is very slow processive microtubule elongation, which recapitulates axonemal dynamics in cells..
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Preprint| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
bioRxiv.org - (2024) vom: 28. März Zur Gesamtaufnahme - year:2024 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Saunders, Harriet A. J. [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2024.03.25.586532 |
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| PPN (Katalog-ID): |
XBI043056237 |
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| 245 | 1 | 0 | |a A network of interacting ciliary tip proteins with opposing activities imparts slow and processive microtubule growth |
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| 520 | |a Abstract Cilia are essential motile or sensory organelles found on many eukaryotic cells. Their formation and function rely on axonemal microtubules, which exhibit very slow dynamics, however the underlying biochemical mechanisms are largely unexplored. Here, we reconstitutedin vitrothe individual and collective activities of the ciliary tip module proteins, CEP104, CSPP1, TOGARAM1, ARMC9 and CCDC66, which interact with each other and with microtubules, and, when mutated, cause ciliopathies such as Joubert syndrome. CEP104, a protein containing a tubulin-binding TOG domain, is an inhibitor of microtubule growth and shortening that interacts with EBs on the microtubule surface and with a luminal microtubule-pausing factor CSPP1. Another TOG-domain protein, TOGARAM1, overcomes growth inhibition imposed by CEP104 and CSPP1. CCDC66 and ARMC9 do not affect microtubule dynamics directly but act as scaffolds for their partners. Cryo-electron tomography showed that together, ciliary tip module members form plus-end-specific cork-like structures which reduce protofilament flaring. The combined effect of these proteins is very slow processive microtubule elongation, which recapitulates axonemal dynamics in cells. | ||
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| 700 | 1 | |a van den Berg, Cyntha M. |0 (orcid)0000-0002-6781-5736 |4 aut | |
| 700 | 1 | |a Hoogebeen, Robin |4 aut | |
| 700 | 1 | |a Schweizer, Donna |4 aut | |
| 700 | 1 | |a Stecker, Kelly E. |0 (orcid)0000-0002-8374-689X |4 aut | |
| 700 | 1 | |a Roepman, Ronald |0 (orcid)0000-0002-5178-8163 |4 aut | |
| 700 | 1 | |a Howes, Stuart C. |0 (orcid)0000-0001-6129-1882 |4 aut | |
| 700 | 1 | |a Akhmanova, Anna |0 (orcid)0000-0002-9048-8614 |4 aut | |
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