Dynamin 2 binds γ-tubulin and participates in centrosome cohesion

Heather M. Thompson, Hong Cao, Jing Chen, Ursula Euteneuer, Mark A Mc Niven

Research output: Contribution to journalArticle

130 Citations (Scopus)

Abstract

Dynamin 2 (Dyn2) is a large GTPase involved in vesicle formation and actin reorganization1-3. In this study, we report a novel role for Dyn2 as a component of the centrosome that is involved in centrosome cohesion. By light microscopy, Dyn2 localized aside centrin and colocalized with γ-tubulin at the centrosome; by immunoelectron microscopy, however, Dyn2 was detected in the pericentriolar material as well as on centrioles. Exogenously expressed green fluorescent protein (GFP)-tagged Dyn2 also localized to the centrosome, whereas glutathione S-transferase (GST)-tagged Dyn2 pulled down a protein complex(es) containing actin, α-tubulin and γ-tubulin from liver homogenate. Furthermore, gel overlay and immunoprecipitation indicated a direct interaction between γ-tubulin and a 219-amino-acid middle domain of Dyn2. Reduction of Dyn2 protein levels with small-interfering RNA (siRNA) resulted in centrosome splitting, whereas microtubule nucleation from centrosomes was not affected, suggesting a role for Dyn2 in centrosome cohesion. Finally, fluorescence recovery after photobleaching (FRAP) analysis of a GFP-tagged Dyn2 middle domain indicated that Dyn2 is a dynamic exchangeable component of the centrosome. These findings suggest a novel function for Dyn2 as a participant in centrosome cohesion.

Original languageEnglish (US)
Pages (from-to)335-342
Number of pages8
JournalNature Cell Biology
Volume6
Issue number4
DOIs
StatePublished - Apr 2004

Fingerprint

Dynamin II
Centrosome
Tubulin
Green Fluorescent Proteins
Actins
Fluorescence Recovery After Photobleaching
Centrioles
Immunoelectron Microscopy
GTP Phosphohydrolases
Sulfamethoxazole Drug Combination Trimethoprim
Glutathione Transferase
Immunoprecipitation

ASJC Scopus subject areas

  • Cell Biology

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Dynamin 2 binds γ-tubulin and participates in centrosome cohesion. / Thompson, Heather M.; Cao, Hong; Chen, Jing; Euteneuer, Ursula; Mc Niven, Mark A.

In: Nature Cell Biology, Vol. 6, No. 4, 04.2004, p. 335-342.

Research output: Contribution to journalArticle

Thompson, Heather M. ; Cao, Hong ; Chen, Jing ; Euteneuer, Ursula ; Mc Niven, Mark A. / Dynamin 2 binds γ-tubulin and participates in centrosome cohesion. In: Nature Cell Biology. 2004 ; Vol. 6, No. 4. pp. 335-342.
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