Removal of the projection domain of microtubule-associated protein 2 alters its interaction with tubulin.
Microtubule-associated proteins (MAPs) can promote microtubule assembly in vitro. One of these MAPs (MAP2) consists of a short promoter domain which binds to the microtubule and promotes assembly and a long projection domain which projects out from the microtubule and may interact with other cytoskeletal elements. We have previously shown that MAP2 and another MAP, tau, differ in their interactions with tubulin in that tau, but not MAP2, promotes extensive aggregation of tubulin into spiral clusters in the presence of vinblastine and that microtuubles formed with MAP2 are more resistant than those formed with tau to the antimitotic drug maytansine [Luduena, R. F., et al. (1984), J. Biol. Chem. 259, 12890-12898; Fellous, A., et al. (1985), Cancer Res. 45, 5004-5010]. Here we have used chymotryptic digestion to remove the projection domain of MAP2 and examined the interaction of the digested MAP2 (ctMAP2) with tubulin in the presence of vinblastine and maytansine. We have found that ctMAP2 behaves very much like tau, but not like undigested MAP2, in the presence of vinblastine, in that ctMAP2 causes tubulin to polymerize into large clusters of spirals. In contrast, microtubule assembly in the presence of ctMAP2 is much more resistant to maytansine inhibition than is assembly in the presence of tau or undigested MAP2. Our results suggest that the projection domain of MAP2 may play a role in the interaction of tubulin with MAP2 during microtubule assembly.
Fellous
A
A
Unité de Recherche sur la Glande Thyroide et la Régulation Hórmonale, INSERM, Bicêtre, France.
Prasad
V
V
Ohayon
R
R
Jordan
M A
MA
Ludueña
R F
RF
D001665:Binding Sites; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008453:Maytansine / Q000494:pharmacology; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D014747:Vinblastine / Q000494:pharmacology; D016875:tau Proteins / Q000737:chemistry
D008869:Microtubule-Associated Proteins; D014404:Tubulin; D016875:tau Proteins; D008453:Maytansine; D014747:Vinblastine