Ca2+/calmodulin-dependent protein kinase IV-mediated LIM kinase activation is critical for calcium signal-induced neurite outgrowth.
Actin cytoskeletal remodeling is essential for neurite outgrowth. LIM kinase 1 (LIMK1) regulates actin cytoskeletal remodeling by phosphorylating and inactivating cofilin, an actin filament-disassembling factor. In this study, we investigated the role of LIMK1 in calcium signal-induced neurite outgrowth. The calcium ionophore ionomycin induced LIMK1 activation and cofilin phosphorylation in Neuro-2a neuroblastoma cells. Knockdown of LIMK1 or expression of a kinase-dead mutant of LIMK1 suppressed ionomycin-induced cofilin phosphorylation and neurite outgrowth in Neuro-2a cells. Ionomycin-induced cofilin phosphorylation and neurite outgrowth were also blocked by KN-93, an inhibitor of Ca(2+)/calmodulin-dependent protein kinases (CaMKs), and STO-609, an inhibitor of CaMK kinase. An active form of CaMKIV but not CaMKI enhanced Thr-508 phosphorylation of LIMK1 and increased the kinase activity of LIMK1. Moreover, the active form of CaMKIV induced cofilin phosphorylation and neurite outgrowth, and a dominant negative form of CaMKIV suppressed ionomycin-induced neurite outgrowth. Taken together, our results suggest that LIMK1-mediated cofilin phosphorylation is critical for ionomycin-induced neurite outgrowth and that CaMKIV mediates ionomycin-induced LIMK1 activation.
Takemura
Miyohiko
M
Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
Mishima
Toshiaki
T
Wang
Yan
Y
Kasahara
Jiro
J
Fukunaga
Kohji
K
Ohashi
Kazumasa
K
Mizuno
Kensaku
K
D000199:Actins / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000737:chemistry; D054734:Calcium-Calmodulin-Dependent Protein Kinase Type 4 / Q000737:chemistry*; D045744:Cell Line, Tumor; D051338:Cofilin 1 / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism; D004789:Enzyme Activation; D015759:Ionomycin / Q000494:pharmacology; D054461:Lim Kinases / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D016501:Neurites / Q000378:metabolism*; D010766:Phosphorylation; D015398:Signal Transduction
D000199:Actins; C495811:Cfl1 protein, mouse; D051338:Cofilin 1; D015759:Ionomycin; D054461:Lim Kinases; D054734:Calcium-Calmodulin-Dependent Protein Kinase Type 4; D002118:Calcium
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