GTP is required for the microtubule catastrophe-inducing activity of MAP200, a tobacco homolog of XMAP215.
Widely conserved among eukaryotes, the microtubule-associated protein 215 (MAP215) family enhances microtubule dynamic instability. The family member studied most extensively, Xenopus laevis XMAP215, has been reported to enhance both assembly and disassembly parameters, although the mechanism whereby one protein can exert these apparently contradictory effects has not been clarified. Here, we analyze the activity of a plant MAP215 homolog, tobacco (Nicotiana tabacum) MAP200 on microtubule behavior in vitro. We show that, like XMAP215, MAP200 promotes both assembly and disassembly parameters, including microtubule growth rate and catastrophe frequency. When MAP200 is added to tubulin and taxol, strikingly long-coiled structures form. When GDP partially replaces GTP, the increase of catastrophe frequency by MAP200 is strongly diminished, even though this replacement stimulates catastrophe in the absence of MAP200. This implies that MAP200 induces catastrophes by a specific, GTP-requiring pathway. We hypothesize that, in the presence of MAP200, a catastrophe-prone microtubule lattice forms occasionally when elongated but nonadjacent protofilaments make lateral contacts.
Hamada
Takahiro
T
Department of Life Science, Graduate School of Life Science, University of Hyogo, Harima Science Park City, Hyogo 678-1297, Japan. hama@bs.naist.jp
Itoh
Tomohiko J
TJ
Hashimoto
Takashi
T
Shimmen
Teruo
T
Sonobe
Seiji
S
D000818:Animals; D006153:Guanosine Diphosphate / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D008869:Microtubule-Associated Proteins / Q000737:chemistry*; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D008954:Models, Biological; D017239:Paclitaxel / Q000494:pharmacology; D010940:Plant Proteins / Q000494:pharmacology* / Q000648:ultrastructure; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000187:drug effects* / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism / Q000648:ultrastructure; D014981:Xenopus; D029867:Xenopus Proteins / Q000737:chemistry*
C486738:CKAP5 protein, Xenopus; D008869:Microtubule-Associated Proteins; D010940:Plant Proteins; D014404:Tubulin; D029867:Xenopus Proteins; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D017239:Paclitaxel