Regulation and subcellular localization of the microtubule-destabilizing stathmin family phosphoproteins in cortical neurons.
Stathmin is a ubiquitous cytosolic phosphoprotein, preferentially expressed in the nervous system, and the generic element of a protein family that includes the neural-specific proteins SCG10, SCLIP, and RB3 and its splice variants, RB3' and RB3". All phosphoproteins of the family share with stathmin its tubulin binding and microtubule (MT)-destabilizing activities. To understand better the specific roles of these proteins in neuronal cells, we performed a comparative study of their expression, regulation, and intracellular distribution in embryonic cortical neurons in culture. We found that stathmin is highly expressed ( approximately 0.25% of total proteins) and uniformly present in the various neuronal compartments (cell body, dendrites, axon, growth cones). It appeared mainly unphosphorylated or weakly phosphorylated on one site, and antisera to specific phosphorylated sites (serines 16, 25, or 38) did not reveal a differential regulation of its phosphorylation among neuronal cell compartments. However, they revealed a subpopulation of cells in which stathmin was highly phosphorylated on serine 16, possibly by CaM kinase II also active in a similar subpopulation. The other proteins of the stathmin family are expressed about 100-fold less than stathmin in partially distinct neuronal populations, RB3 being detected in only about 20% of neurons in culture. In contrast to stathmin, they are each mostly concentrated at the Golgi apparatus and are also present along dendrites and axons, including growth cones. Altogether, our results suggest that the different members of the stathmin family have complementary, at least partially distinct functions in neuronal cell regulation, in particular in relation to MT dynamics.
INSERM U440, Institut du Fer à Moulin, Paris, France.
Olivier
O
Gavet
Saïd
S
El Messari
Sylvie
S
Ozon
André
A
Sobel
D000818:Animals; D000918:Antibody Specificity; D001369:Axons / Q000737:chemistry; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D003712:Dendrites / Q000737:chemistry; D006056:Golgi Apparatus / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D051379:Mice; D008868:Microtubule Proteins; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D010750:Phosphoproteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D050777:Stathmin
D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008868:Microtubule Proteins; D009414:Nerve Growth Factors; D010750:Phosphoproteins; D050777:Stathmin; C493994:Stmn1 protein, mouse; C512849:Stmn2 protein, mouse; C512848:Stmn2 protein, rat; C491793:Stmn3 protein, mouse; C491792:Stmn3 protein, rat; C491794:Stmn4 protein, mouse; C112286:Stmn4 protein, rat