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Article Dans Une Revue Journal of Cell Biology Année : 2021

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

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
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Etsuko Muto
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Résumé

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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Dates et versions

hal-03423264 , version 1 (10-11-2021)

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Rie Ayukawa, Seigo Iwata, Hiroshi Imai, Shinji Kamimura, Masahito Hayashi, et al.. GTP-dependent formation of straight tubulin oligomers leads to microtubule nucleation. Journal of Cell Biology, 2021, 220, ⟨10.1083/jcb.202007033⟩. ⟨hal-03423264⟩
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