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Published Online October 16, 2008
Science DOI: 10.1126/science.1165401

Research Articles

Submitted on September 3, 2008
Accepted on October 8, 2008

Detection of GTP-Tubulin Conformation in Vivo Reveals a Role for GTP Remnants in Microtubule Rescues

Ariane Dimitrov 1{dagger}, Mélanie Quesnoit 2{dagger}, Sandrine Moutel 3, Isabelle Cantaloube 4, Christian Poüs 5{ddagger}, Franck Perez 1{ddagger}*

1 CNRS UMR144, Institut Curie, 26 rue d’Ulm, 75248 Paris Cedex 05, France.; Institut Curie, Centre de Recherche, 26 rue d’Ulm, 75248 Paris Cedex 05, France.
2 CNRS UMR144, Institut Curie, 26 rue d’Ulm, 75248 Paris Cedex 05, France.; Institut Curie, Centre de Recherche, 26 rue d’Ulm, 75248 Paris Cedex 05, France.; Biochimie et Biologie Cellulaire–JE2493, Université Paris-Sud 11, IFR141–Faculté de Pharmacie, Châtenay-Malabry, France.
3 Institut Curie, Centre de Recherche, 26 rue d’Ulm, 75248 Paris Cedex 05, France.; Institut Curie, Translational Research Department, 26 rue d’Ulm, 75248 Paris Cedex 05, France.
4 Biochimie et Biologie Cellulaire–JE2493, Université Paris-Sud 11, IFR141–Faculté de Pharmacie, Châtenay-Malabry, France.
5 Biochimie et Biologie Cellulaire–JE2493, Université Paris-Sud 11, IFR141–Faculté de Pharmacie, Châtenay-Malabry, France.; Biochimie-Hormonologie, AP-HP, Hôpital Antoine Béclère, Clamart, France.

* To whom correspondence should be addressed.
Franck Perez , E-mail: franck.perez{at}curie.fr

{dagger}These authors contributed equally to this work.

{ddagger}These authors contributed equally to this work.

Microtubules display dynamic instability, with alternating phases of growth and shrinkage separated by catastrophe and rescue events. The GTP cap at the growing end of microtubules is essential to prevent microtubule catastrophes in vitro, but has been difficult to observe in vivo. Here, we selected a recombinant antibody specifically recognizing GTP-bound tubulin in microtubules and found that GTP-tubulin was, indeed, present at the plus end of growing microtubules. Unexpectedly, GTP-tubulin remnants were also present in older parts of microtubules suggesting that GTP hydrolysis is sometimes incomplete during polymerization. Observations in living cells suggested that these GTP-remnants may be responsible for the rescue events in which microtubules recover from catastrophe.


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