GTP-dependent formation of straight tubulin oligomers leads to microtubule nucleation | Zendy

Rie Ayukawa | Zendy; Seigo Iwata | Zendy; Hiroshi Imai | Zendy; Shinji Kamimura | Zendy; Masahito Hayashi | Zendy; Kien Xuan Ngo | Zendy; Itsushi Minoura | Zendy; Seiichi Uchimura | Zendy; Tsukasa Makino | Zendy; Mikako Shirouzu | Zendy; Hideki Shigematsu | Zendy; Ken Sekimoto | Zendy; Benoı̂t Gigant | Zendy; Etsuko Muto | Zendy

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GTP-dependent formation of straight tubulin oligomers leads to microtubule nucleation

Author(s) -

Rie Ayukawa,

Seigo Iwata,

Hiroshi Imai,

Shinji Kamimura,

Masahito Hayashi,

Kien Xuan Ngo,

Itsushi Minoura,

Seiichi Uchimura,

Tsukasa Makino,

Mikako Shirouzu,

Hideki Shigematsu,

Ken Sekimoto,

Benoı̂t Gigant,

Etsuko Muto

Publication year - 2021

Publication title -

the journal of cell biology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 5.414

H-Index - 380

eISSN - 1540-8140

pISSN - 0021-9525

DOI - 10.1083/jcb.202007033

Subject(s) - nucleation , tubulin , gtp' , microtubule , microtubule nucleation , biophysics , chemistry , oligomer , negative stain , population , crystallography , biology , electron microscope , biochemistry , cell , polymer chemistry , microbiology and biotechnology , cell cycle , centrosome , physics , enzyme , organic chemistry , demography , sociology , optics

Nucleation of microtubules (MTs) is essential for cellular activities, but its mechanism is unknown because of the difficulty involved in capturing rare stochastic events in the early stage of polymerization. Here, combining rapid flush negative stain electron microscopy (EM) and kinetic analysis, we demonstrate that the formation of straight oligomers of critical size is essential for nucleation. Both GDP and GTP tubulin form single-stranded oligomers with a broad range of curvatures, but upon nucleation, the curvature distribution of GTP oligomers is shifted to produce a minor population of straight oligomers. With tubulin having the Y222F mutation in the β subunit, the proportion of straight oligomers increases and nucleation accelerates. Our results support a model in which GTP binding generates a minor population of straight oligomers compatible with lateral association and further growth to MTs. This study suggests that cellular factors involved in nucleation promote it via stabilization of straight oligomers.

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