Glutamylated tubulin probed in ciliates with the monoclonal antibody GT335.
Microtubular networks are extensively developed in many ciliate species. In several of them, we investigate the occurrence of the post-translational glutamylation of tubulin [Eddé et al., 1990: Science 247:82-85; Eddé et al., 1991: J. Cell. Biochem. 46:134-142] using as a probe for such modified tubulin, the monoclonal antibody GT335 [Wolff et al., 1992: Eur. J. Cell Biol. 59:425-432]. Results obtained in Paramecium strongly suggest that both axonemal and cytoplasmic tubulin are glutamylated. As in the vertebrate brain tubulin so far tested, the GT335 epitope is located at the carboxy-terminal fragment of cytoplasmic tubulin removed by subtilisin treatment. Immunoblotting and immunofluorescence experiments reveal that, unlike tubulin acetylation, glutamylation is not restricted to cold-resistant microtubules. In addition, immunofluorescence studies performed on dividing cells show that glutamylation takes place soon after the polymerization of microtubules. Finally, glutamylated tubulin is also detected in the ciliate species Euplotes, Tetrahymena, and Paraurostyla. Together with results obtained on flagellate species, this suggests that tubulin glutamylation came out early in the course of eukaryotic evolution and has been widely exploited in various cellular strategies.
Bré
M H
MH
Laboratoire de Biologie Cellulaire 4, CNRS URA 1134, Université Paris XI, Orsay, France.
de Néchaud
B
B
Wolff
A
A
Fleury
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D002923:Cilia / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry; D000939:Epitopes; D056890:Eukaryota / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure; D005971:Glutamates; D018698:Glutamic Acid; D008870:Microtubules / Q000737:chemistry; D015335:Molecular Probes; D010247:Paramecium / Q000276:immunology; D014404:Tubulin / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005971:Glutamates; D015335:Molecular Probes; D014404:Tubulin; D018698:Glutamic Acid