Plant Physiol. Drug Metab Dispos
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Published on October 23, 2009; 10.1104/pp.109.144303

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Received July 5, 2009
Accepted October 21, 2009

GTP is required for the microtubule catastrophe-inducing activity of MAP200, a tobacco homolog of XMAP215

Takahiro Hamada *, Tomohiko J. Itoh , Takashi Hashimoto , Teruo Shimmen , and Seiji Sonobe

Department of Life Science, Graduate School of Life Science, University of Hyogo, Harima Science Park City, Hyogo, 678-1297, Japan; Division of Biological Sciences, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan; Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630–0101, Japan

* Corresponding author; email: hama{at}bs.naist.jp.

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 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 non-adjacent protofilaments make lateral contacts.






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