TY - JOUR
T1 - Actin filaments regulate microtubule growth at the centrosome
AU - Inoue, Daisuke
AU - Obino, Dorian
AU - Pineau, Judith
AU - Farina, Francesca
AU - Gaillard, Jérémie
AU - Guerin, Christophe
AU - Blanchoin, Laurent
AU - Lennon-Duménil, Ana Maria
AU - Théry, Manuel
N1 - Funding Information:
DI was supported by a fellowship from the Uehara Memorial Foundation (Japan). DO was supported by a fellowship from the Fondation pour la Recherche Médicale (FDT20150532056). Funding was obtained from the Association Nationale pour la Recherche (ANR-PoLyBex-12-BSV3-0014-001 to A-ML-D and ANR-Mitotube-12-BSV5-0004-01 to MT) and the European Research Council (ERC-Strapacemi-GA 243103 to A-ML-D and ERC-SpiCy 310472 to MT).
Funding Information:
We acknowledge the Recombinant Antibodies Platform (TAb-IP) at Institut Curie, Paris, France for providing the VHH anti-GFP-human Fc antibody. We thank Pablo Saez (Institut Curie, France) for providing the EB3-mCherry plasmid and Robert Robinson (IMCB, Singapore) for providing the purified gelsolin. DI was supported by a fellowship from the Uehara Memorial Foundation (Japan). DO was supported by a fellowship from the Fondation pour la Recherche M?dicale (FDT20150532056). Funding was obtained from the Association Nationale pour la Recherche (ANR-PoLyBex-12-BSV3-0014-001 to A-ML-D and ANR-Mitotube-12-BSV5-0004-01 to MT) and the European Research Council (ERC-Strapacemi-GA 243103 to A-ML-D and ERC-SpiCy 310472 to MT).
Publisher Copyright:
© 2019 The Authors
PY - 2019/6/3
Y1 - 2019/6/3
N2 - The centrosome is the main microtubule-organizing centre. It also organizes a local network of actin filaments. However, the precise function of the actin network at the centrosome is not well understood. Here, we show that increasing densities of actin filaments at the centrosome of lymphocytes are correlated with reduced amounts of microtubules. Furthermore, lymphocyte activation resulted in disassembly of centrosomal actin and an increase in microtubule number. To further investigate the direct crosstalk between actin and microtubules at the centrosome, we performed in vitro reconstitution assays based on (i) purified centrosomes and (ii) on the co-micropatterning of microtubule seeds and actin filaments. These two assays demonstrated that actin filaments constitute a physical barrier blocking elongation of nascent microtubules. Finally, we showed that cell adhesion and cell spreading lead to lower densities of centrosomal actin, thus resulting in higher microtubule growth. We therefore propose a novel mechanism, by which the number of centrosomal microtubules is regulated by cell adhesion and actin-network architecture.
AB - The centrosome is the main microtubule-organizing centre. It also organizes a local network of actin filaments. However, the precise function of the actin network at the centrosome is not well understood. Here, we show that increasing densities of actin filaments at the centrosome of lymphocytes are correlated with reduced amounts of microtubules. Furthermore, lymphocyte activation resulted in disassembly of centrosomal actin and an increase in microtubule number. To further investigate the direct crosstalk between actin and microtubules at the centrosome, we performed in vitro reconstitution assays based on (i) purified centrosomes and (ii) on the co-micropatterning of microtubule seeds and actin filaments. These two assays demonstrated that actin filaments constitute a physical barrier blocking elongation of nascent microtubules. Finally, we showed that cell adhesion and cell spreading lead to lower densities of centrosomal actin, thus resulting in higher microtubule growth. We therefore propose a novel mechanism, by which the number of centrosomal microtubules is regulated by cell adhesion and actin-network architecture.
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U2 - 10.15252/embj.201899630
DO - 10.15252/embj.201899630
M3 - Article
C2 - 30902847
AN - SCOPUS:85063276813
SN - 0261-4189
VL - 38
JO - EMBO Journal
JF - EMBO Journal
IS - 11
M1 - e99630
ER -