Abstract
Mitogen-activated protein kinase (MAPK)-activated protein kinase 2 (MK2) is activated upon stress by p38 MAPK alpha and -beta, which bind to a basic docking motif in the C terminus of MK2 and which subsequently phosphorylate its regulatory sites. As a result of activation MK2 is exported from the nucleus to the cytoplasm and cotransports active p38 MAPK to this compartment. Here we show that the amount of p38 MAPK is significantly reduced in cells and tissues lacking MK2, indicating a stabilizing effect of MK2 for p38. Using a murine knockout model, we have previously shown that elimination of MK2 leads to a dramatic reduction of tumor necrosis factor (TNF) production in response to lipopolysaccharide. To further elucidate the role of MK2 in p38 MAPK stabilization and in TNF biosynthesis, we analyzed the ability of two MK2 isoforms and several MK2 mutants to restore both p38 MAPK protein levels and TNF biosynthesis in macrophages. We show that MK2 stabilizes p38 MAPK through its C terminus and that MK2 catalytic activity does not contribute to this stabilization. Importantly, we demonstrate that stabilizing p38 MAPK does not restore TNF biosynthesis. TNF biosynthesis is only restored with MK2 catalytic activity. We further show that, in MK2-deficient macrophages, formation of filopodia in response to extracellular stimuli is reduced. In addition, migration of MK2-deficient mouse embryonic fibroblasts (MEFs) and smooth muscle cells on fibronectin is dramatically reduced. Interestingly, reintroducing catalytic MK2 activity into MEFs alone is not sufficient to revert the migratory phenotype of these cells. In addition to catalytic activity, the proline-rich N-terminal region is necessary for rescuing the migratory phenotype. These data indicate that catalytic activity of MK2 is required for both cytokine production and cell migration. However, the proline-rich MK2 N terminus provides a distinct role restricted to cell migration.
MeSH Terms
Adenoviridae/genetics
Amino Acid Motifs
Amino Acid Sequence
Animals
Cell Movement/genetics
Cells, Cultured
Fibroblasts
Intracellular Signaling Peptides and Proteins
Lipopolysaccharides/pharmacology
Macrophages/drug effects,metabolism
Mice
Mice, Mutant Strains
Mitogen-Activated Protein Kinases/genetics,metabolism
Molecular Sequence Data
Muscle, Smooth/cytology
Mutation
Nuclear Localization Signals/genetics,metabolism
Organ Specificity
Proline
Protein Kinases
Protein Serine-Threonine Kinases
Protein Structure, Tertiary
Tumor Necrosis Factor-alpha/metabolism
p38 Mitogen-Activated Protein Kinases
Chemicals
Intracellular Signaling Peptides and Proteins
Lipopolysaccharides
Nuclear Localization Signals
Tumor Necrosis Factor-alpha
Proline
Protein Kinases
MAP-kinase-activated kinase 2
Protein Serine-Threonine Kinases
Mitogen-Activated Protein Kinases
p38 Mitogen-Activated Protein Kinases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Kotlyarov Alexey
Institute of Biochemistry, Medical School Hannover, Hannover 30625, Germany.
Yannoni Yvonne
Fritz Susann
Laass Kathrin
Telliez Jean-Baptiste
Pitman Deborah
Lin Lih-Ling
Gaestel Matthias
References (22)
22 references, click to expand
-
The Drosophila melanogaster homolog of the mammalian MAPK-activated protein kinase-2 (MAPKAPK-2) lacks a proline-rich N-terminus.
Gene. 1995 Oct 3;163(2):209-14
PMID: 7590268
-
Pro-inflammatory cytokines and environmental stress cause p38 mitogen-activated protein kinase activation by dual phosphorylation on tyrosine and threonine.
J Biol Chem. 1995 Mar 31;270(13):7420-6
PMID: 7535770
-
Identification of novel phosphorylation sites required for activation of MAPKAP kinase-2.
EMBO J. 1995 Dec 1;14(23):5920-30
PMID: 8846784
-
Identification of mitogen-activated protein (MAP) kinase-activated protein kinase-3, a novel substrate of CSBP p38 MAP kinase.
J Biol Chem. 1996 Apr 5;271(14):8488-92
PMID: 8626550
-
Identification of MAPKAPK homolog (MAPKAPK-4) as a myosin II regulatory light-chain kinase in sea urchin egg extracts.
Arch Biochem Biophys. 1997 Jul 1;343(1):55-62
PMID: 9210646
-
PMA-induced activation of the p42/44ERK- and p38RK-MAP kinase cascades in HL-60 cells is PKC dependent but not essential for differentiation to the macrophage-like phenotype.
J Cell Physiol. 1997 Dec;173(3):310-8
PMID: 9369943
-
Leptomycin B-sensitive nuclear export of MAPKAP kinase 2 is regulated by phosphorylation.
EMBO J. 1998 Jun 15;17(12):3363-71
PMID: 9628873
-
PRAK, a novel protein kinase regulated by the p38 MAP kinase.
EMBO J. 1998 Jun 15;17(12):3372-84
PMID: 9628874
-
Nuclear export of the stress-activated protein kinase p38 mediated by its substrate MAPKAP kinase-2.
Curr Biol. 1998 Sep 24;8(19):1049-57
PMID: 9768359
-
A role for p38(MAPK)/HSP27 pathway in smooth muscle cell migration.
J Biol Chem. 1999 Aug 20;274(34):24211-9
PMID: 10446196
-
MAPKAP kinase 2 is essential for LPS-induced TNF-alpha biosynthesis.
Nat Cell Biol. 1999 Jun;1(2):94-7
PMID: 10559880
-
The role of thin filopodia in motility and morphogenesis.
Exp Cell Res. 1999 Dec 15;253(2):296-301
PMID: 10585250
-
A conserved docking motif in MAP kinases common to substrates, activators and regulators.
Nat Cell Biol. 2000 Feb;2(2):110-6
PMID: 10655591
-
Identification of a docking groove on ERK and p38 MAP kinases that regulates the specificity of docking interactions.
EMBO J. 2001 Feb 1;20(3):466-79
PMID: 11157753
-
alpha 2 integrin subunit cytoplasmic domain-dependent cellular migration requires p38 MAPK.
J Biol Chem. 2001 Mar 23;276(12):9503-11
PMID: 11121402
-
Identification of MAPKAP kinase 2 as a major enzyme responsible for the phosphorylation of the small mammalian heat shock proteins.
FEBS Lett. 1992 Nov 30;313(3):307-13
PMID: 1332886
-
Modulation of actin microfilament dynamics and fluid phase pinocytosis by phosphorylation of heat shock protein 27.
J Biol Chem. 1993 Nov 15;268(32):24210-4
PMID: 8226968
-
The substrate specificity and structure of mitogen-activated protein (MAP) kinase-activated protein kinase-2.
Biochem J. 1993 Dec 15;296 ( Pt 3):843-9
PMID: 8280084
-
The primary structure of a human MAP kinase activated protein kinase 2.
Biochem Biophys Res Commun. 1994 Apr 29;200(2):1118-24
PMID: 8179591
-
Phosphorylation and supramolecular organization of murine small heat shock protein HSP25 abolish its actin polymerization-inhibiting activity.
J Biol Chem. 1994 Aug 12;269(32):20780-4
PMID: 8051180
-
A protein kinase involved in the regulation of inflammatory cytokine biosynthesis.
Nature. 1994 Dec 22-29;372(6508):739-46
PMID: 7997261
-
Constitutive activation of mitogen-activated protein kinase-activated protein kinase 2 by mutation of phosphorylation sites and an A-helix motif.
J Biol Chem. 1995 Nov 10;270(45):27213-21
PMID: 7592979