Differential effect of two stathmin/Op18 phosphorylation mutants on Xenopus embryo development.
Stathmin/Op18 destabilizes microtubules in vitro and regulates microtubule polymerization in vivo. Both a microtubule catastrophe-promoting activity and a tubulin sequestering activity were demonstrated for stathmin in vitro, and both could contribute to microtubule depolymerization in vivo. Stathmin activity can be turned down by extensive phosphorylation on its four phosphorylatable serines, and down-regulation of stathmin activity by phosphorylation is necessary for cells to proceed through mitosis. We show here that microinjection of a nonphosphorylatable Ser to Ala (4A) quadruple mutant in Xenopus two-cell stage embryos results in cell cleavage arrest in the injected blastomeres and aborted development, whereas injection of a pseudo-phosphorylated Ser to Glu quadruple mutant (4E) does not prevent normal development. Addition of these mutants to mitotic cytostatic factor-arrested extracts in which spindle assembly was induced led to a dramatic reduction of spindle size with 4A stathmin, and to a moderate increase with 4E stathmin, but both localized to spindle poles. Interestingly, the microtubule assembly-dependent phosphorylation of endogenous stathmin was abolished in the presence of 4A stathmin, but not of 4E stathmin. Altogether, this shows that the phosphorylation-mediated regulation of stathmin activity during the cell cycle is essential for early Xenopus embryonic development.
Küntziger
T
T
Institut Curie, Section Recherche, UMR 144 CNRS, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
Gavet
O
O
Sobel
A
A
Bornens
M
M
D000818:Animals; D004625:Embryo, Nonmammalian / Q000378:metabolism; D047108:Embryonic Development; D006801:Humans; D008856:Microscopy, Fluorescence; D008868:Microtubule Proteins; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D050777:Stathmin; D014981:Xenopus / Q000196:embryology; D029867:Xenopus Proteins
D008868:Microtubule Proteins; D010750:Phosphoproteins; C493996:STMN1 protein, Xenopus; C493995:STMN1 protein, human; D050777:Stathmin; D029867:Xenopus Proteins