MARCKS dephosphorylation is involved in bradykinin-induced neurite outgrowth in neuroblastoma SH-SY5Y cells

Atsuhiro Tanabe, Mitsuya Shiraishi, Manabu Negishi, Naoaki Saito, Mitsuo Tanabe, Yasuharu Sasaki

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Bradykinin (BK) plays a major role in producing peripheral sensitization in response to peripheral inflammation and in pain transmission in the central nerve system (CNS). Because BK activates protein kinase C (PKC) through phospholipase C (PLC)-β and myristoylated alanine-rich C kinase substrate (MARCKS) has been found to be a substrate of PKC, we explored the possibility that BK could induce MARCKS phosphorylation and regulate its function. BK stimulation induced transient MARCKS phosphorylation on Ser159 with a peak at 1min in human neuroblastoma SH-SY5Y cells. By contrast, PKC activation by the phorbol ester phorbol 12,13-dibutyrate (PDBu) elicited MARCKS phosphorylation which lasted more than 10min. Western blotting analyses and glutathione S-transferase (GST) pull-down analyses showed that the phosphorylation by BK was the result of activation of the PKC-dependent RhoA/Rho-associated coiled-coil kinase (ROCK) pathway. Protein phosphatase (PP) 2A inhibitors calyculin A and fostriecin inhibited the dephosphorylation of MARCKS after BK-induced phosphorylation. Moreover, immunoprecipitation analyses showed that PP2A interacts with MARCKS. These results indicated that PP2A is the dominant PP of MARCKS after BK stimulation. We established SH-SY5Y cell lines expressing wild-type MARCKS and unphosphorylatable MARCKS, and cell morphology changes after cell stimulation were studied. PDBu induced lamellipodia formation on the neuroblastoma cell line SH-SY5Y and the morphology was sustained, whereas BK induced neurite outgrowth of the cells via lamellipodia-like actin accumulation that depended on transient MARCKS phosphorylation. Thus these findings show a novel BK signal cascade-that is, BK promotes neurite outgrowth through transient MARCKS phosphorylation involving the PKC-dependent RhoA/ROCK pathway and PP2A in a neuroblastoma cell line.

Original languageEnglish
Pages (from-to)618-629
Number of pages12
JournalJournal of Cellular Physiology
Volume227
Issue number2
DOIs
Publication statusPublished - 2012 Jan
Externally publishedYes

Fingerprint

Bradykinin
Phosphorylation
Protein Kinase C
Phorbol 12,13-Dibutyrate
rho-Associated Kinases
Cells
myristoylated alanine-rich C kinase substrate
Chemical activation
Protein Phosphatase 2
Phosphoprotein Phosphatases
Type C Phospholipases
Phorbol Esters
Glutathione Transferase
Actins

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

MARCKS dephosphorylation is involved in bradykinin-induced neurite outgrowth in neuroblastoma SH-SY5Y cells. / Tanabe, Atsuhiro; Shiraishi, Mitsuya; Negishi, Manabu; Saito, Naoaki; Tanabe, Mitsuo; Sasaki, Yasuharu.

In: Journal of Cellular Physiology, Vol. 227, No. 2, 01.2012, p. 618-629.

Research output: Contribution to journalArticle

Tanabe, Atsuhiro ; Shiraishi, Mitsuya ; Negishi, Manabu ; Saito, Naoaki ; Tanabe, Mitsuo ; Sasaki, Yasuharu. / MARCKS dephosphorylation is involved in bradykinin-induced neurite outgrowth in neuroblastoma SH-SY5Y cells. In: Journal of Cellular Physiology. 2012 ; Vol. 227, No. 2. pp. 618-629.
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AU - Tanabe, Atsuhiro

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AU - Saito, Naoaki

AU - Tanabe, Mitsuo

AU - Sasaki, Yasuharu

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