Activation of specific MEK-ERK cascade is necessary for TGF beta signalingand crosstalk with PKA and PKC pathways in cultured rat articular chondrocytes
Y. Hirota et al., Activation of specific MEK-ERK cascade is necessary for TGF beta signalingand crosstalk with PKA and PKC pathways in cultured rat articular chondrocytes, OSTEO CART, 8(4), 2000, pp. 241-247
Objective: TGF beta is a potent stimulator of cell growth in cultured rat a
rticular chondrocytes (CRAC). The stimulatory effect is mediated through th
e immediate induction of c-fos gene by activating ERK of MAPK. The present
study was undertaken to investigate the upstream regulators involved in TGF
beta-induced ERK activation in CRAC and to compare the results with the ev
ents in HepG2 cells.
Results: in vitro kinase and trans-reporting assays showed that TGF beta pr
eferentially activated ERK and JNK pathways in CRAC and HepG2, respectively
. ERK activation in CRAC was selectively inhibited by PD98059, a MEK inhibi
tor. Overexpression of wild or active forms of MEKK1, the upstream activato
r of ERK and JNK, decreased the TGF beta-induced 3TP-luciferase activity in
CRAC. In contrast, in HepG2 dominant negative form of MEKK1 or SEK1 ligand
-dependent reporter activity was diminished. Transfection of TAK1, another
MAPKKK, also positively and negatively regulated 3TP transcriptional activi
ty of HepG2 and CRAC, respectively Activation of PKA by 8-bromo-cyclic AMP
or forskolin, and inhibition of PKC by calphostin C, resulted in a signific
ant decrease in 3TP activity as well as in vitro ERK kinase activity in CRA
C.
Conclusions: The results indicate that TGF beta transduces a predominant si
gnal pathway through MEK-ERK-Elk1, independent of MEKK1 or TAK1 pathway in
CRAC. However, in HepG2, activation of MEKK1 and TAK1 is essential for TGF
beta-induced signal transmission. The results also demonstrated that in CRA
C, MEK-ERK pathway activated by TGF beta is negatively regulated by PKA cas
cade but transactivated by PKC. (C) 2000 OsteoArthritis Research Society In
ternational.