Phosphorylation of the Kinase Interaction Motif in Mitogen-activated Protein (MAP) Kinase Phosphatase-4 Mediates Cross-talk between Protein Kinase A and MAP Kinase Signaling Pathways

Robin J. Dickinson, Laurent Delavaine, Rocio Cejudo-Marin, Graeme Stewart, Christopher J. Staples, Mark P. Didmon, Antonio Garcia Trinidad, Andres Alonso, Rafael Pulido, Stephen M. Keyse (Lead / Corresponding author)

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    14 Citations (Scopus)

    Abstract

    MAP kinase phosphatase 4 (DUSP9/MKP-4) plays an essential role during placental development and is one of a subfamily of three closely related cytoplasmic dual-specificity MAPK phosphatases, which includes the ERK-specific enzymes DUSP6/MKP-3 and DUSP7/MKP-X. However, unlike DUSP6/MKP-3, DUSP9/MKP-4 also inactivates the p38 alpha MAP kinase both in vitro and in vivo. Here we demonstrate that inactivation of both ERK1/2 and p38 alpha by DUSP9/MKP-4 is mediated by a conserved arginine-rich kinase interaction motif located within the amino-terminal non-catalytic domain of the protein. Furthermore, DUSP9/MKP-4 is unique among these cytoplasmic MKPs in containing a conserved PKA consensus phosphorylation site 55RRXSer-58 immediately adjacent to the kinase interaction motif. DUSP9/MKP-4 is phosphorylated on Ser-58 by PKA in vitro, and phosphorylation abrogates the binding of DUSP9/MKP-4 to both ERK2 and p38 alpha MAP kinases. In addition, although mutation of Ser-58 to either alanine or glutamic acid does not affect the intrinsic catalytic activity of DUSP9/MKP-4, phospho-mimetic (Ser-58 to Glu) substitution inhibits both the interaction of DUSP9/MKP-4 with ERK2 and p38 alpha in vivo and its ability to dephosphorylate and inactivate these MAP kinases. Finally, the use of a phospho-specific antibody demonstrates that endogenous DUSP9/MKP-4 is phosphorylated on Ser-58 in response to the PKA agonist forskolin and is also modified in placental tissue. We conclude that DUSP9/MKP-4 is a bona fide target of PKA signaling and that attenuation of DUSP9/MKP-4 function can mediate cross-talk between the PKA pathway and MAPK signaling through both ERK1/2 and p38 alpha in vivo.

    Original languageEnglish
    Pages (from-to)38018-38026
    Number of pages9
    JournalJournal of Biological Chemistry
    Volume286
    Issue number44
    DOIs
    Publication statusPublished - 4 Nov 2011

    Keywords

    • DUAL-SPECIFICITY PHOSPHATASES
    • TYROSINE-PHOSPHATASE
    • CATALYTIC ACTIVATION
    • DIFFERENTIAL REGULATION
    • INSULIN-RESISTANCE
    • PTP-SL
    • CAMP
    • ERK2
    • INACTIVATION
    • ASSOCIATION

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