Exogenous C2-ceramide activates c-fos serum response element via Rac-dependent signalling pathway

Byung Chul Kim, Jae-Hong Kim

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Ceramide is an important regulatory molecule implicated in a variety of biological processes in response to stress and cytokines. To understand the signal transduction pathway of ceramide to the nucleus, in the present study, we examined whether C2-ceramide a cell permeable ceramide, activates c-fos serum response element (SRE). Treatment of Rat-2 fibroblast cells with C2-ceramide caused the stimulation of c-fos SRE-dependent reporter gene activity in a dose- and time-dependent manner by transient transfection analysis. Next, we examined the role of Rho family GTPases in the ceramide-induced signalling to SRE activation. By reporter gene analysis following transient transfections with various plasmids expressing a dominant negative mutant form of Cdc42, Rac1 or RhoA, C2-ceramide-induced SRE activation was shown to be selectively repressed by pEXV-RacN17 encoding a dominant negative mutant of Rac1, suggesting that Rac activity is essential for the signalling cascade of ceramide to the nucleus. In a further study to analyse the downstream mediator of Rac in the ceramide-signalling pathway, we observed that either pretreatment with mepacrine, a potent and specific inhibitor of phospholipase A2, or co-transfection with antisense cytosolic phospholipase A2 (cPLA2) oligonucleotide repressed the C2-ceramide-induced SRE activation selectively, implying a critical role of cPLA2 in C2-ceramide-induced signalling to nucleus. Consistent with these results, the translocation of cPLA2 protein as well as the release of arachidonic acid, a principal product of phospholipase A2, was rapidly induced by the addition of C2-ceramide in a Rac-dependent manner. Together, our findings suggest the critical role of 'Rac and subsequent activation of phospholipase A2' in ceramide-signalling to nucleus.

Original languageEnglish
Pages (from-to)1009-1014
Number of pages6
JournalBiochemical Journal
Volume330
Issue number2
Publication statusPublished - 1998 Mar 1
Externally publishedYes

Fingerprint

Serum Response Element
Ceramides
Cytosolic Phospholipases A2
Chemical activation
Transfection
Phospholipases A2
Reporter Genes
Transient analysis
Genes
Biological Phenomena
Quinacrine
Signal transduction
rho GTP-Binding Proteins
GTP Phosphohydrolases
Fibroblasts
N-acetylsphingosine
Arachidonic Acid
Oligonucleotides
Rats
Signal Transduction

ASJC Scopus subject areas

  • Biochemistry

Cite this

Exogenous C2-ceramide activates c-fos serum response element via Rac-dependent signalling pathway. / Kim, Byung Chul; Kim, Jae-Hong.

In: Biochemical Journal, Vol. 330, No. 2, 01.03.1998, p. 1009-1014.

Research output: Contribution to journalArticle

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