TY - JOUR
T1 - PKC phosphorylates MARCKS Ser159 not only directly but also through RhoA/ROCK
AU - Tanabe, Atsuhiro
AU - Kamisuki, Yukinori
AU - Hidaka, Hiroyoshi
AU - Suzuki, Masaaki
AU - Negishi, Manabu
AU - Takuwa, Yoh
N1 - Funding Information:
This work was supported in part by Grants-in-Aid for Scientific Research (16290069) from the Japan Society for the Promotion of Science, by grants from the Takeda Science Foundation, and by a Kitasato University Research Grant for Young Researchers.
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2006/6/23
Y1 - 2006/6/23
N2 - It is well recognized that phorbol 12,13-dibutyrate (PDBu)-activated PKC directly phosphorylates myristoylated alanine-rich C kinase substrate (MARCKS), whose phosphorylation is used as a marker of PKC activation. However, in SH-SY5Y neuroblastoma cells, Western blotting analyses revealed that Rho-associated coiled-coil kinase (ROCK)-specific inhibitor H-1152 inhibited PDBu-induced phosphorylation, and that a small G-protein inhibitor, toxin B, also inhibited MARCKS phosphorylation. Furthermore, in GST pull-down assays, PDBu induced RhoA activation in SH-SY5Y cells, and this activation was inhibited by PKC inhibitor Ro-31-8220. Finally, we showed that the transfection of a dominant negative form of RhoA inhibited PDBu-induced MARCKS phosphorylation in immunocytochemistries. These findings suggest that some PDBu-induced MARCKS phosphorylation includes the RhoA/ROCK pathway in SH-SY5Y cells.
AB - It is well recognized that phorbol 12,13-dibutyrate (PDBu)-activated PKC directly phosphorylates myristoylated alanine-rich C kinase substrate (MARCKS), whose phosphorylation is used as a marker of PKC activation. However, in SH-SY5Y neuroblastoma cells, Western blotting analyses revealed that Rho-associated coiled-coil kinase (ROCK)-specific inhibitor H-1152 inhibited PDBu-induced phosphorylation, and that a small G-protein inhibitor, toxin B, also inhibited MARCKS phosphorylation. Furthermore, in GST pull-down assays, PDBu induced RhoA activation in SH-SY5Y cells, and this activation was inhibited by PKC inhibitor Ro-31-8220. Finally, we showed that the transfection of a dominant negative form of RhoA inhibited PDBu-induced MARCKS phosphorylation in immunocytochemistries. These findings suggest that some PDBu-induced MARCKS phosphorylation includes the RhoA/ROCK pathway in SH-SY5Y cells.
KW - MARCKS
KW - Neuroblastoma
KW - PKC
KW - Phosphorylation
KW - ROCK
KW - RhoA
UR - http://www.scopus.com/inward/record.url?scp=33646401566&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33646401566&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2006.04.082
DO - 10.1016/j.bbrc.2006.04.082
M3 - Article
C2 - 16677610
AN - SCOPUS:33646401566
SN - 0006-291X
VL - 345
SP - 156
EP - 161
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -