Microtubule-associated proteins control the kinetics of microtubule nucleation

Nat Cell Biol. 2015 Jul;17(7):907-16. doi: 10.1038/ncb3188. Epub 2015 Jun 22.


Microtubules are born and reborn continuously, even during quiescence. These polymers are nucleated from templates, namely γ-tubulin ring complexes (γ-TuRCs) and severed microtubule ends. Using single-molecule biophysics, we show that nucleation from γ-TuRCs, axonemes and seed microtubules requires tubulin concentrations that lie well above the critical concentration. We measured considerable time lags between the arrival of tubulin and the onset of steady-state elongation. Microtubule-associated proteins (MAPs) alter these time lags. Catastrophe factors (MCAK and EB1) inhibited nucleation, whereas a polymerase (XMAP215) and an anti-catastrophe factor (TPX2) promoted nucleation. We observed similar phenomena in cells. We conclude that GTP hydrolysis inhibits microtubule nucleation by destabilizing the nascent plus ends required for persistent elongation. Our results explain how MAPs establish the spatial and temporal profile of microtubule nucleation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axoneme / metabolism
  • CHO Cells
  • Cell Line, Tumor
  • Centrosome / metabolism
  • Cricetinae
  • Cricetulus
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Guanosine Triphosphate / analogs & derivatives
  • Guanosine Triphosphate / metabolism
  • Humans
  • Hydrolysis
  • Immunoblotting
  • Kinetics
  • LLC-PK1 Cells
  • Microscopy, Electron
  • Microscopy, Fluorescence / methods
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / drug effects
  • Microtubules / metabolism*
  • Microtubules / ultrastructure
  • Nocodazole / pharmacology
  • Polymerization / drug effects
  • Swine
  • Tubulin / metabolism*
  • Tubulin Modulators / pharmacology


  • Microtubule-Associated Proteins
  • Tubulin
  • Tubulin Modulators
  • Green Fluorescent Proteins
  • 5'-guanylylmethylenebisphosphonate
  • Guanosine Triphosphate
  • Nocodazole