Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends

Saishree S. Iyer; Fangrui Chen; Funso E. Ogunmolu; Shoeib Moradi; Vladimir A. Volkov; Emma J. van Grinsven; Chris van Hoorn; Jingchao Wu; Nemo Andrea; Shasha Hua; Kai Jiang; Ioannis Vakonakis; Mia Potočnjak; Franz Herzog; Benoît Gigant; Nikita Gudimchuk; Kelly E. Stecker; Marileen Dogterom; Michel O. Steinmetz; Anna Akhmanova

doi:10.1083/jcb.202406061

Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends

Saishree S. Iyer, Fangrui Chen, Funso E. Ogunmolu, Shoeib Moradi, Vladimir A. Volkov, Emma J. van Grinsven, Chris van Hoorn, Jingchao Wu, Nemo Andrea, Shasha Hua, Kai Jiang, Ioannis Vakonakis, Mia Potočnjak, Franz Herzog, Benoît Gigant, Nikita Gudimchuk, Kelly E. Stecker, Marileen Dogterom^*, Michel O. Steinmetz^*, Anna Akhmanova^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Centrioles are microtubule-based organelles required for the formation of centrosomes and cilia. Centriolar microtubules, unlike their cytosolic counterparts, are stable and grow very slowly, but the underlying mechanisms are poorly understood. Here, we reconstituted in vitro the interplay between the proteins that cap distal centriole ends and control their elongation: CP110, CEP97, and CPAP/SAS-4. We found that whereas CEP97 does not bind to microtubules directly, CP110 autonomously binds microtubule plus ends, blocks their growth, and inhibits depolymerization. Cryo-electron tomography revealed that CP110 associates with the luminal side of microtubule plus ends and suppresses protofilament flaring. CP110 directly interacts with CPAP, which acts as a microtubule polymerase that overcomes CP110-induced growth inhibition. Together, the two proteins impose extremely slow processive microtubule growth. Disruption of CP110-CPAP interaction in cells inhibits centriole elongation and increases incidence of centriole defects. Our findings reveal how two centriolar cap proteins with opposing activities regulate microtubule plus-end elongation and explain their antagonistic relationship during centriole formation.

Original language	English
Article number	e202406061
Number of pages	36
Journal	The Journal of cell biology
Volume	224
Issue number	3
DOIs	https://doi.org/10.1083/jcb.202406061
Publication status	Published - 3 Mar 2025

Bibliographical note

Publisher Copyright:
© 2025 Iyer et al.

Funding

We thank R. Stucchi and A.F.M. Altelaar (Utrecht University) for the help with mass spectrometry, W. Evers and A. Jakobi lab (TU Delft) for the help with cryo-ET. J.R. McIntosh (University of Colorado) for the feedback on PF shape analysis, and D. Mastronarde (University of Colorado) for modifying the Howflared program to extract 3D coordinates from PF models. We thank the Diamond Light Source for the provision of synchrotron radiation facilities and the staff of Beamline B21 for their help.This work was supported by the European Research Council Synergy grant 609822 to M. Dogterom and A. Akhmanova, the Netherlands Organization for Scientific Research ALWOP.440 grant to A. Akhmanova, the China Scholarship Council scholarship to F. Chen. The Medical Research Council UK (MR/N009274/1) grant to I. Vakonakis, the European Molecular Biology Organization Long Term Fellowship ALTF-840-2018 to F.E. Ogunmolu, the Marie Curie COFUND Fellowship (Marie Sklodowska-Curie grant agreement 701647) to S. Moradi, and the grants from the Swiss National Science Foundation (31003A_166608 and 310030_192566 to M.O. Steinmetz and 310030M_215014 to A. Akhmanova). Analysis of the shapes of tubulin PFs was supported by the Russian Science Foundation grant 21-74-20035 to N. Gudimchuk.

Funders	Funder number
European Research Council	609822
Netherlands Organization for Scientific Research	ALWOP.440
China Scholarship Council scholarship
Medical Research Council UK	MR/N009274/1
European Molecular Biology Organization Long Term Fellowship	ALTF-840-2018
Marie Curie COFUND Fellowship	701647
Swiss National Science Foundation	31003A_166608, 310030_192566, 310030M_215014
Russian Science Foundation	21-74-20035
Swiss National Science Foundation (SNF)	310030_192566

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Iyer, S. S., Chen, F., Ogunmolu, F. E., Moradi, S., Volkov, V. A., van Grinsven, E. J., van Hoorn, C., Wu, J., Andrea, N., Hua, S., Jiang, K., Vakonakis, I., Potočnjak, M., Herzog, F., Gigant, B., Gudimchuk, N., Stecker, K. E., Dogterom, M., Steinmetz, M. O., & Akhmanova, A. (2025). Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends. The Journal of cell biology, 224(3), Article e202406061. https://doi.org/10.1083/jcb.202406061

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title = "Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends",

abstract = "Centrioles are microtubule-based organelles required for the formation of centrosomes and cilia. Centriolar microtubules, unlike their cytosolic counterparts, are stable and grow very slowly, but the underlying mechanisms are poorly understood. Here, we reconstituted in vitro the interplay between the proteins that cap distal centriole ends and control their elongation: CP110, CEP97, and CPAP/SAS-4. We found that whereas CEP97 does not bind to microtubules directly, CP110 autonomously binds microtubule plus ends, blocks their growth, and inhibits depolymerization. Cryo-electron tomography revealed that CP110 associates with the luminal side of microtubule plus ends and suppresses protofilament flaring. CP110 directly interacts with CPAP, which acts as a microtubule polymerase that overcomes CP110-induced growth inhibition. Together, the two proteins impose extremely slow processive microtubule growth. Disruption of CP110-CPAP interaction in cells inhibits centriole elongation and increases incidence of centriole defects. Our findings reveal how two centriolar cap proteins with opposing activities regulate microtubule plus-end elongation and explain their antagonistic relationship during centriole formation.",

author = "Iyer, {Saishree S.} and Fangrui Chen and Ogunmolu, {Funso E.} and Shoeib Moradi and Volkov, {Vladimir A.} and {van Grinsven}, {Emma J.} and {van Hoorn}, Chris and Jingchao Wu and Nemo Andrea and Shasha Hua and Kai Jiang and Ioannis Vakonakis and Mia Poto{\v c}njak and Franz Herzog and Beno{\^i}t Gigant and Nikita Gudimchuk and Stecker, {Kelly E.} and Marileen Dogterom and Steinmetz, {Michel O.} and Anna Akhmanova",

note = "Publisher Copyright: {\textcopyright} 2025 Iyer et al.",

year = "2025",

month = mar,

day = "3",

doi = "10.1083/jcb.202406061",

language = "English",

volume = "224",

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Iyer, SS, Chen, F, Ogunmolu, FE, Moradi, S, Volkov, VA, van Grinsven, EJ, van Hoorn, C, Wu, J, Andrea, N, Hua, S, Jiang, K, Vakonakis, I, Potočnjak, M, Herzog, F, Gigant, B, Gudimchuk, N, Stecker, KE, Dogterom, M, Steinmetz, MO & Akhmanova, A 2025, 'Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends', The Journal of cell biology, vol. 224, no. 3, e202406061. https://doi.org/10.1083/jcb.202406061

TY - JOUR

T1 - Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends

AU - Iyer, Saishree S.

AU - Chen, Fangrui

AU - Ogunmolu, Funso E.

AU - Moradi, Shoeib

AU - Volkov, Vladimir A.

AU - van Grinsven, Emma J.

AU - van Hoorn, Chris

AU - Wu, Jingchao

AU - Andrea, Nemo

AU - Hua, Shasha

AU - Jiang, Kai

AU - Vakonakis, Ioannis

AU - Potočnjak, Mia

AU - Herzog, Franz

AU - Gigant, Benoît

AU - Gudimchuk, Nikita

AU - Stecker, Kelly E.

AU - Dogterom, Marileen

AU - Steinmetz, Michel O.

AU - Akhmanova, Anna

PY - 2025/3/3

Y1 - 2025/3/3

N2 - Centrioles are microtubule-based organelles required for the formation of centrosomes and cilia. Centriolar microtubules, unlike their cytosolic counterparts, are stable and grow very slowly, but the underlying mechanisms are poorly understood. Here, we reconstituted in vitro the interplay between the proteins that cap distal centriole ends and control their elongation: CP110, CEP97, and CPAP/SAS-4. We found that whereas CEP97 does not bind to microtubules directly, CP110 autonomously binds microtubule plus ends, blocks their growth, and inhibits depolymerization. Cryo-electron tomography revealed that CP110 associates with the luminal side of microtubule plus ends and suppresses protofilament flaring. CP110 directly interacts with CPAP, which acts as a microtubule polymerase that overcomes CP110-induced growth inhibition. Together, the two proteins impose extremely slow processive microtubule growth. Disruption of CP110-CPAP interaction in cells inhibits centriole elongation and increases incidence of centriole defects. Our findings reveal how two centriolar cap proteins with opposing activities regulate microtubule plus-end elongation and explain their antagonistic relationship during centriole formation.

AB - Centrioles are microtubule-based organelles required for the formation of centrosomes and cilia. Centriolar microtubules, unlike their cytosolic counterparts, are stable and grow very slowly, but the underlying mechanisms are poorly understood. Here, we reconstituted in vitro the interplay between the proteins that cap distal centriole ends and control their elongation: CP110, CEP97, and CPAP/SAS-4. We found that whereas CEP97 does not bind to microtubules directly, CP110 autonomously binds microtubule plus ends, blocks their growth, and inhibits depolymerization. Cryo-electron tomography revealed that CP110 associates with the luminal side of microtubule plus ends and suppresses protofilament flaring. CP110 directly interacts with CPAP, which acts as a microtubule polymerase that overcomes CP110-induced growth inhibition. Together, the two proteins impose extremely slow processive microtubule growth. Disruption of CP110-CPAP interaction in cells inhibits centriole elongation and increases incidence of centriole defects. Our findings reveal how two centriolar cap proteins with opposing activities regulate microtubule plus-end elongation and explain their antagonistic relationship during centriole formation.

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U2 - 10.1083/jcb.202406061

DO - 10.1083/jcb.202406061

M3 - Article

C2 - 39847124

AN - SCOPUS:85217274207

SN - 0021-9525

VL - 224

JO - The Journal of cell biology

JF - The Journal of cell biology

IS - 3

M1 - e202406061

ER -

Centriolar cap proteins CP110 and CPAP control slow elongation of microtubule plus ends

Abstract

Bibliographical note

Funding

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