Differential distribution of glutamylated tubulin isoforms along the sea urchin sperm axoneme.
Tubulin belongs to a highly conserved multigenic family, in which several gene products usually coexist in the same tissue or the same cell. Moreover, seven classes of post-translational modifications of these gene products lead to an amazing diversity of tubulin polypeptide chains, within the same cell type, whose physiological function remains elusive. Such diversity has been found in a very stable microtubular organelle, the sea urchin sperm flagellum, where some tubulin isoforms have been directly implicated in motility, whereas others may play a more structural role. In particular, polyglutamylated tubulin has been shown to be crucial for motility (Gagnon et al., 1996: J Cell Sci 109:1545 p). Here, we show with the GT335 antibody that polyglutamylated tubulin is distributed according to a decreasing gradient along the sea urchin sperm axoneme, since a semi-quantitative measurement of immunofluorescence intensity reveals that in its proximal half, the axoneme is sixfold more labeled than in its distal half. This gradient along the length of the axoneme is confirmed by immunogold labeling procedures which, in addition, demonstrate a uniform distribution of polyglutamylated tubulin among peripheral doublets and a lesser content in the central pair within a same section. Moreover, our data obtained with B3, an antibody that recognizes both mono- and poly-glutamylated tubulin, suggest that the number of glutamate residues in the lateral poly-glutamyl chain of tubulin varies along the whole length of the axoneme. These novel results coupled with those published earlier may be important to understand the role of polyglutamylation in flagellar motility.
Huitorel
Philippe
P
Laboratoire de Biologie du Développement, UMR 7009 CNRS et Univ. P. & M. Curie, Station Zoologique, Observatoire Océanologique, 06234 Villefranche-sur-Mer, France. huitorel@obs-vlfr.fr
White
Daniel
D
Fouquet
Jean-Pierre
JP
Kann
Marie-Louise
ML
Cosson
Jacky
J
Gagnon
Claude
C
D000818:Animals; D005455:Fluorescent Antibody Technique; D018698:Glutamic Acid / Q000378:metabolism*; D008297:Male; D016253:Microscopy, Immunoelectron; D020033:Protein Isoforms; D012617:Sea Urchins; D013094:Spermatozoa / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*
D020033:Protein Isoforms; D014404:Tubulin; D018698:Glutamic Acid