Drosophila stathmins bind tubulin heterodimers with high and variable stoichiometries.
In vertebrates, stathmins form a family of proteins possessing two tubulin binding repeats (TBRs), which each binds one soluble tubulin heterodimer. The stathmins thus sequester two tubulins in a phosphorylation-dependent manner, providing a link between signal transduction and microtubule dynamics. In Drosophila, we show here that a single stathmin gene (stai) encodes a family of D-stathmin proteins. Two of the D-stathmins are maternally deposited and then restricted to germ cells, and the other two are detected in the nervous system during embryo development. Like in vertebrates, the nervous system-enriched stathmins contain an N-terminal domain involved in subcellular targeting. All the D-stathmins possess a domain containing three or four predicted TBRs, and we demonstrate here, using complementary biochemical and biophysical methods, that all four predicted TBR domains actually bind tubulin. D-stathmins can indeed bind up to four tubulins, the resulting complex being directly visualized by electron microscopy. Phylogenetic analysis shows that the presence of regulated multiple tubulin sites is a conserved characteristic of stathmins in invertebrates and allows us to predict key residues in stathmin for the binding of tubulin. Altogether, our results reveal that the single Drosophila stathmin gene codes for a stathmin family similar to the multigene vertebrate one, but with particular tubulin binding properties.
Lachkar
Sylvie
S
INSERM U 839, Paris F-75005, France.
Lebois
Marion
M
Steinmetz
Michel O
MO
Guichet
Antoine
A
Lal
Neha
N
Curmi
Patrick A
PA
Sobel
André
A
Ozon
Sylvie
S
D000818:Animals; D019281:Dimerization; D004330:Drosophila; D006367:HeLa Cells; D006801:Humans; D017403:In Situ Hybridization; D008870:Microtubules / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D011994:Recombinant Proteins / Q000737:chemistry; D050777:Stathmin / Q000737:chemistry* / Q000235:genetics*; D020349:Surface Plasmon Resonance; D014404:Tubulin / Q000737:chemistry*
D011994:Recombinant Proteins; D050777:Stathmin; D014404:Tubulin