Negative-feedback regulation of FGF signalling by DUSP6/MKP-3 is driven by ERK1/2 and mediated by Ets factor binding to a conserved site within the DUSP6/MKP-3 gene promoter

Abstract

DUSP6 (dual-specificity phosphatase 6), also known as MKP-3 [MAPK (mitogen-activated protein kinase) phosphatase-3] specifically inactivates ERK1/2 (extracellular-signal-regulated kinase 1/2) in vitro and in vivo. DUSP6/MKP-3 is inducible by FGF (fibroblast growth factor) signalling and acts as a negative regulator of ERK activity in key and discrete signalling centres that direct outgrowth and patterning in early vertebrate embryos. However, the molecular mechanism by which FGFs induce DUSP6/MKP-3 expression and hence help to set ERK1/2 signalling levels is unknown. In the present study, we demonstrate, using pharmacological inhibitors and analysis of the murine DUSP6/MKP-3 gene promoter, that the ERK pathway is critical for FGF-induced DUSP6/MKP-3 transcription. Furthermore, we show that this response is mediated by a conserved binding site for the Ets (E twenty-six) family of transcriptional regulators and that the Ets2 protein, a known target of ERK signalling, binds to the endogenous DUSP6/MKP-3 promoter. Finally, the murine DUSP6/MKP-3 promoter coupled to EGFP (enhanced green fluorescent protein) recapitulates the specific pattern of endogenous DUSP6/MKP-3 mRNA expression in the chicken neural plate, where its activity depends on FGFR (FGF receptor) and MAPK signalling and an intact Ets-binding site. These findings identify a conserved Ets-factor-dependent mechanism by which ERK signalling activates DUSP6/MKP-3 transcription to deliver ERK1/2-specific negative-feedback control of FGF signalling.

Follicle-Stimulating Hormone (FSH)-dependent Regulation of Extracellular Regulated Kinase (ERK) Phosphorylation by the Mitogen-activated Protein (MAP) Kinase Phosphatase MKP3

“…Regulation of MKP3(DUPS6) expression is most often placed downstream of ERK signaling (63,64) and is thought to constitute a negative feedback loop to downregulate mitogenic signaling. Consistent with this conclusion, Mkp3/Dusp6 is often induced following ERK activation (65) and requires regulation by the transcription factor Ets1 (64,65). Upon expression, MKP3 is catalytically activated via its physical association with ERK (62,66,67).…”

DUSP1/MKP1 promotes angiogenesis, invasion and metastasis in non-small-cell lung cancer

“…As the sequence of DUSP6 promoter has recently been reported (Ekerot et al, 2008;Morrison et al, 2008), we investigated if DUSP1 was able to modulate DUSP6 expression by stimulating the promoter activity. Co-transfection of a DUSP6 promoter reporter vector (Prom DUSP6) and CTAPDUSP1, a strong and transient DUSP1 expression vector in 293T and H460 cells, induced the activation of DUSP6 by threefold and 1.5-fold, respectively (Figure 1d).…”

“…Although it has been reported that ERK activity modulates DUSP6 expression in response to fibroblast growth factor (Ekerot et al, 2008), in H460 cells expression of DUSP1 increases both DUSP6 promoter activation and expression, thus maybe other kinases, such as p38a and/or JNK, should control DUSP6 expression by repressing its promoter. DUSP6 can be upregulated in specific cancers exhibiting aberrant RTK and ras/raf signaling such as NSCLC (Sato et al, 2006).…”

Cisplatin regulates the MAPK kinase pathway to induce increased expression of DNA repair gene ERCC1 and increase melanoma chemoresistance

“…However, another cause of ERK activation can be inhibition of ERK phosphatases, such as DUSP6 (Kondoh and Nishida, 2007). Both mathematical models and experimental data showed that DUSP6 was the principal time regulator of ERK signalling (Ekerot et al, 2008). Thus, we speculated that the cisplatin-mediated, delayed and long-lasting activation of ERK was, at least in part, because of inhibition of the ERK-specific phosphatase, DUSP6.…”