MAP65 in tubulin/colchicine paracrystals of Vigna sinensis root cells: possible role in the assembly and stabilization of atypical tubulin polymers.
Members of the MAP65 family, colocalizing with microtubule arrays, have been identified in Vigna sinensis root cells by Western blotting and immunofluorescence. MAP65 proteins were also found in tubulin/colchicine paracrystals, which were formed during colchicine treatment by both immunofluorescence and immunogold microscopy. During recovery from colchicine, MAP65 signal was depleted from disintegrating paracrystals appearing in the reinstating microtubule arrays. MAP65-free perinuclear tubulin/colchicine aggregates were observed in plasmolyzed colchicine-treated cells. Deplasmolysis of the above cells resulted in the formation of MAP65-decorated paracrystals. As confirmed by appropriate biochemical assays with the Phos-tag reagent, MAP65 proteins underwent phosphorylation during plasmolysis, which was reversible by deplasmolysis. According to the effect of the mitogen-activated protein kinase (MAPK) inhibitor UO126, the phosphorylation status of MAP65, as well as its presence in tubulin/colchicine polymers is probably controlled by MAPK-mediated phosphorylation. According to the above, it seems likely that apart from binding to microtubules, MAP65 proteins may act as "tubulin associated proteins" in a broader manner, promoting the polymerization and/or stabilization of atypical polymers such as tubulin/colchicine paracrystals.
Panteris
Emmanuel
E
Department of Botany, School of Biology, Aristotle University, Thessaloniki, Macedonia, Greece. epanter@bio.auth.gr
Komis
George
G
Adamakis
Ioannis-Dimosthenis S
ID
Samaj
Jozef
J
Bosabalidis
Artemios M
AM
D003078:Colchicine / Q000378:metabolism / Q000494:pharmacology*; D003460:Crystallization; D007887:Fabaceae / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D018517:Plant Roots / Q000187:drug effects / Q000378:metabolism; D020033:Protein Isoforms / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
C514153:1,3-bis(bis(pyridin-2-ylmethyl)amino)propan-2-ol; D008869:Microtubule-Associated Proteins; D020033:Protein Isoforms; D011725:Pyridines; D014404:Tubulin; D003078:Colchicine