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MBC in Press, published online ahead of print February 4, 2002
Mol. Biol. Cell 10.1091/mbc.01-06-0298

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Submitted on June 20, 2001
Revised on November 1, 2001
Accepted on December 13, 2001

Two related sub-pellicular cytoskeleton-associated proteins in Trypanosoma brucei stabilise microtubules

Cécile Vedrenne1, Christiane Giroud1, Derrick R. Robinson2, Sébastien Besteiro1, Christophe Bosc3, Frédéric Bringaud1*, and Théo Baltz1

1 Laboratoire de Parasitologie Moléculaire, Université Victor Segalen de Bordeaux II, UMR-5016 CNRS, 146 rue Léo Saignat, 33076 Bordeaux cedex, France
2 University of Manchester, Stopford Bldg, 2.205 Oxford road, Manchester, M13 9PT, England, U.K. (present address: Laboratoire de Parasitologie Moléculaire, Université Victor Segalen de Bordeaux II, UMR-5016 CNRS, 146 rue Léo Saignat, 33076 Bordeaux cedex, France)
3 Commissariat à l'Energie Atomique, Laboratoire du Cytosquelette, Institut National de la Santé et de la Recherche Médicale Unité 366, 17 rue des Martyrs, 38054 Grenoble cedex 9, France

* Corresponding author. E-mail address: bringaud{at}u-bordeaux2.fr.

The sub-pellicular microtubules of the trypanosome cytoskeleton are cross-linked to each other and the plasma membrane creating a cage-like structure. We have isolated from Trypanosoma brucei, two related low molecular-weight proteins (15 and 17 kDa), called CAP15 and CAP17, which are differentialy expressed during the life cycle. Immunolabelling shows a corset-like co-localisation of both anti-CAP antisera similar to that of anti-tubulin labelling. Western blot and electron microscope analyses show CAP15 and CAP17 is present on detergent extracted cytoskeletons. However, the localisation of both proteins is restricted to the anterior, microtubule minus and less dynamic half of the corset. CAP15 and CAP17 share properties of microtubule-associated proteins when expressed in heterologous cells, (Chinese hamster ovary and HeLa) co-localisation with microtubules, induction of microtubule bundle formation, cold-resistance and insensitivity to nocodazole. When over-expressed in T. brucei, both CAP15 and CAP17 cover the whole sub-pellicular corset and induce morphological disorders, cell cycle based abnormalities and subsequent asymetric cytokinesis.




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