Mitogen-activated-protein-kinase-catalyzed phosphorylation of microtubule-associated proteins, microtubule-associated protein 2 and microtubule-associated protein 4, induces an alteration in their function.
Mitogen-activated protein kinase (MAPK), a serine/threonine-specific protein kinase which is generally activated by stimulation with various growth factors and phorbol esters, utilizes microtubule-associated protein (MAP) 2 as a good substrate in vitro. We have found that MAPK-catalyzed phosphorylation of MAP2 resulted in a significant loss in its ability to induce tubulin polymerization. The chymotryptic fragments, containing a microtubule-binding domain of MAP2, were phosphorylated by MAPK and the ability of the fragments to induce tubulin polymerization was also greatly decreased by the phosphorylation, suggesting that phosphorylation of the microtubule-binding domain is important for functional alteration of MAP2. In addition to MAP2, a 190-kDa heat-stable MAP (MAP4) found in various tissues and cells, was a good substrate for MAPK in vitro. Phosphorylation of MAP4 inactivated tubulin polymerization. We examined the effect of phosphorylation of MAP2 and MAP4 on the dynamics of microtubules nucleated by purified centrosomes in vitro. The data showed that MAPK-catalyzed phosphorylation of MAP2 and MAP4 reduced their ability to increase the apparent elongation rate and the number of microtubules nucleated by the centrosome. Thus, MAPK is capable of phosphorylating MAPs and negatively regulating their microtubule-stabilizing function.
Hoshi
M
M
Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
Ohta
K
K
Gotoh
Y
Y
Mori
A
A
Murofushi
H
H
Sakai
H
H
Nishida
E
E
D000818:Animals; D001345:Autoradiography; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002384:Catalysis; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005455:Fluorescent Antibody Technique; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008934:Mitogens / Q000494:pharmacology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D013552:Swine; D014404:Tubulin / Q000378:metabolism
D008869:Microtubule-Associated Proteins; D008934:Mitogens; D014404:Tubulin; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases