Dynamic clustering of dynamin-amphiphysin helices regulates membrane constriction and fission coupled with GTP hydrolysis

Tetsuya Takeda, Toshiya Kozai, Huiran Yang, Daiki Ishikuro, Kaho Seyama, Yusuke Kumagai, Tadashi Abe, Hiroshi Yamada, Takayuki Uchihashi, Toshio Ando, Kohji Takei

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Dynamin is a mechanochemical GTPase essential for membrane fission during clathrin-mediated endocytosis. Dynamin forms helical complexes at the neck of clathrin-coated pits and their structural changes coupled with GTP hydrolysis drive membrane fission. Dynamin and its binding protein amphiphysin cooperatively regulate membrane remodeling during the fission, but its precise mechanism remains elusive. In this study, we analyzed structural changes of dynamin-amphiphysin complexes during the membrane fission using electron microscopy (EM) and high-speed atomic force microscopy (HS-AFM). Interestingly, HS-AFM analyses show that the dynamin-amphiphysin helices are rearranged to form clusters upon GTP hydrolysis and membrane constriction occurs at protein-uncoated regions flanking the clusters. We also show a novel function of amphiphysin in size control of the clusters to enhance biogenesis of endocytic vesicles. Our approaches using combination of EM and HS-AFM clearly demonstrate new mechanistic insights into the dynamics of dynamin-amphiphysin complexes during membrane fission.

Original languageEnglish
Article numbere30246
JournaleLife
Volume7
DOIs
Publication statusPublished - Jan 23 2018

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Dynamins
Guanosine Triphosphate
Constriction
Cluster Analysis
Hydrolysis
Membranes
Atomic Force Microscopy
Atomic force microscopy
Clathrin
Electron microscopy
Electron Microscopy
Transport Vesicles
GTP Phosphohydrolases
Endocytosis
amphiphysin
Carrier Proteins
Neck

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Dynamic clustering of dynamin-amphiphysin helices regulates membrane constriction and fission coupled with GTP hydrolysis. / Takeda, Tetsuya; Kozai, Toshiya; Yang, Huiran; Ishikuro, Daiki; Seyama, Kaho; Kumagai, Yusuke; Abe, Tadashi; Yamada, Hiroshi; Uchihashi, Takayuki; Ando, Toshio; Takei, Kohji.

In: eLife, Vol. 7, e30246, 23.01.2018.

Research output: Contribution to journalArticle

Takeda, Tetsuya ; Kozai, Toshiya ; Yang, Huiran ; Ishikuro, Daiki ; Seyama, Kaho ; Kumagai, Yusuke ; Abe, Tadashi ; Yamada, Hiroshi ; Uchihashi, Takayuki ; Ando, Toshio ; Takei, Kohji. / Dynamic clustering of dynamin-amphiphysin helices regulates membrane constriction and fission coupled with GTP hydrolysis. In: eLife. 2018 ; Vol. 7.
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AU - Ishikuro, Daiki

AU - Seyama, Kaho

AU - Kumagai, Yusuke

AU - Abe, Tadashi

AU - Yamada, Hiroshi

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AU - Ando, Toshio

AU - Takei, Kohji

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