The effect of stathmin phosphorylation on microtubule assembly depends on tubulin critical concentration.
Stathmin is a phosphorylation-regulated tubulin-binding protein. In vitro and in vivo studies using nonphosphorylatable and pseudophosphorylated mutants of stathmin have questioned the view that stathmin might act only as a tubulin-sequestering factor. Stathmin was proposed to effectively regulate microtubule dynamic instability by increasing the frequency of catastrophe (the transition from steady growth to rapid depolymerization), without interacting with tubulin. We have used a noninvasive method to measure the equilibrium dissociation constants of the T(2)S complexes of tubulin with stathmin, pseudophosphorylated (4E)-stathmin, and diphosphostathmin. At both pH 6.8 and pH 7.4, the relative sequestering efficiency of the different stathmin variants depends on the concentration of free tubulin, i.e. on the dynamic state of microtubules. This control is exerted in a narrow range of tubulin concentration due to the highly cooperative binding of tubulin to stathmin. Changes in pH affect the stability of tubulin-stathmin complexes but do not change stathmin function. The 4E-stathmin mutant mimics inactive phosphorylated stathmin at low tubulin concentration and sequesters tubulin almost as efficiently as stathmin at higher tubulin concentration. We propose that stathmin acts solely by sequestering tubulin, without affecting microtubule dynamics, and that the effect of stathmin phosphorylation on microtubule assembly depends on tubulin critical concentration.
Amayed
Phedra
P
Dynamique du Cytosquelette, Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, 91198 Gif-sur-Yvette, France.
Pantaloni
Dominique
D
Carlier
Marie-France
MF
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002850:Chromatography, Gel; D000077143:Docetaxel; D004305:Dose-Response Relationship, Drug; D006863:Hydrogen-Ion Concentration; D008868:Microtubule Proteins; D008870:Microtubules / Q000378:metabolism*; D008956:Models, Chemical; D017239:Paclitaxel / Q000031:analogs & derivatives* / Q000494:pharmacology; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D050777:Stathmin; D043823:Taxoids; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
D000972:Antineoplastic Agents, Phytogenic; D008868:Microtubule Proteins; D010750:Phosphoproteins; D050777:Stathmin; D043823:Taxoids; D014404:Tubulin; D000077143:Docetaxel; D017239:Paclitaxel