Trp-Lys-Tyr-Met-Val-Met activates mitogen-activated protein kinase via a PI-3 kinase-mediated pathway independent of PKC

doi:10.1016/S0024-3205(99)00436-1

Life Sciences

Volume 65, Issue 17, 17 September 1999, Pages 1845-1856

https://doi.org/10.1016/S0024-3205(99)00436-1 Get rights and content

Abstract

Trp-Lys-Tyr-Met-Val-Met (WKYMVM) is a novel potent peptide which can stimulate phosphoinositide hydrolysis in U937 as well as U266 and HL-60 cells (Baek et al., J. Biol. Chem. 271, 8170 (1996)). The peptide also induces Superoxide generation in human neutrophils (Seo et al., J. Immunol. 158, 1896 (1997)). However, the signaling pathway down-stream of PLC set in motion by the peptide is not yet completely understood. We studied the signaling pathway of the peptide with the goal of elucidating the mechanism of the peptide's action. WKYMVM induced a rapid and transient activation of the ERKs in human histiocytic lymphoma cells, U937. The ERK1 activation peaked at 5 min and returned to the basal level after 30 min. The ERK1 stimulation by the peptide was partially inhibited by pretreatment of the cells with pertussis toxin (PTX), implicating G-protein involvement in the peptide's action. Pretreatment of staurosporine, protein kinase C (PKC) inhibitor, or PKC down-regulating PMA had no impact on the ERK1 activation by the peptide, indicating that the signaling pathway is independent of PKC activation. Pretreatment of the cells with neomycin and intracellular Ca²⁺ mobilizing reagents had also no effect on the ERK1 activation by the peptide. However, pretreatment with wortmannin or LY294002, the inhibitor of phosphatidylinositol 3 kinase (PI-3K), strongly inhibited peptide-stimulated ERK1 activation. Our results suggest that PI-3K may be an important participant in the ERK cascade induced by the peptide. Furthermore, the treatment of U937 cells with the peptide activated p74^raf-1, an upstream kinase of ERK. Taken together, our results suggest that the peptide activate ERK via a G-protein/PI-3K/Ras/Raf-1 mediated signaling pathway in U937 cells.

References (43)

H. Yu et al.
J. Biol. Chem.
(1995)
M.H. Pillinger et al.
J. Biol. Chem.
(1996)
A. Ptasznik et al.
J. Biol. Chem.
(1996)
Y.L. Zu et al.
Blood
(1996)
J. Troppmair et al.
J. Biol. Chem.
(1994)
C.J. Marshall
Cell
(1995)
S. Grinstein et al.
J. Biol. Chem.
(1992)
C. Knall et al.
J. Biol. Chem.
(1996)
S. Grinstein et al.
J. Biol. Chem.
(1994)
A.M. Buhl et al.
J. Biol. Chem.
(1995)

S.H. Baek et al.

J. Biol. Chem.

(1996)

J.K. Seo et al.

Clin. Biochem.

(1998)

EM Walker et al.

Biochem. Pharmacol.

(1998)

M. Ohmichi et al.

J. Biol. Chem.

(1994)

T. Okada et al.

J. Biol. Chem.

(1994)

M. Thelen et al.

Biochem. Biophys. Res. Commun.

(1995)

N.J. Avdi et al.

J. Biol Chem.

(1996)

I.M. Ferby et al.

J. Biol Chem.

(1994)

K. Yamauchi et al.

J. Biol. Chem.

(1993)

A. Sjolander et al.

Biochem. Biophys. Res. Commun.

(1992)

M.B. Hallet

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Inhibitory analysis demonstrated that fMLF-stimulated ERK activation depends on tyrosine kinases, PI3K, PKC, and PLC [57]. Upon stimulation of U937 cells with WKYMVM ERK phosphorylation was observed after 5 min, partially suppressed by pertussis toxin and significantly by phosphatidylinositol-3-kinase inhibitors, but did not depend on PKC and Ca2+mobilization [58]. Both WKYMVM and LXA4 were shown to cause chemotaxis, an increase in [Ca2+]i, and PKC activation, but only pro-inflammatory peptide agonists activated MAPK, cPLA2, and NADPH oxidase-dependent ROS production in human neutrophils [59,60].
Formyl peptide receptors (FPRs) are expressed in the cells of the innate immune system and provide binding with pathogen and damage-associated molecular patterns with subsequent activation of the phagocytes for defense reactions such as chemotaxis, secretory degranulation and ROS generation. Probably, FPR2 is one of the unique receptors in the organism; it is able to recognize numerous ligands of different chemical structure, and moreover, these ligands can trigger opposite phagocyte responses promoting either pro- or anti-inflammatory reactions. Therefore, FPR2 and its signaling pathways are of intense research interest.
We found only slight activation of ERK1/2 in the response to peptide ligand WKYMVM in the accelerating phase of ROS generation and more intense ERK1/2 phosphorylation in the declining phase of it in mouse bone marrow granulocytes. Lipid agonist BML-111 did not induce significant ERK phosphorylation when applied for 10–1800 s.
To some extent co-localization of ERK1/2 and NADPH oxidase subunits was observed even in the intact cells and didn't change under FPR2 stimulation by WKYMVM, while direct PKC activation by PMA resulted to more efficient interaction between ERK1/2 and p47phox/p67phox and their translocation to plasma membrane. We have shown that phosphorylation and activation of ERK1/2 in bone marrow granulocytes depended on FPR2-triggered activity of PI3K and PKC, phosphatase DUSP6, and, the most but not the least, on ROS generation. Since blocking of ROS generation led to a slowdown of ERK activation indicating a significant contribution of ROS to the secondary regulation of ERK activity.
Formyl peptide receptor like 1 differentially requires mitogen-activated protein kinases for the induction of glial fibrillary acidic protein and interleukin-1α in human U87 astrocytoma cells
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Citation Excerpt :
The initial suggestion for the participation of PI3K in the pathway from GPCR toward MAPK cascades came from several reports that specific inhibitors of PI3K, wortmannin and LY294002, strongly abolished ERK and JNK activation induced by LPA, α2-adrenergic receptor or Gβγ subunits [50,31]. Similarly, studies in human U937 lymphoma cells and human monocytes have found that WKYMVM and WKYMVm evoked a PI3K-dependent, but PKC-independent ERK activation [11,51]. Nevertheless, our experiments did not support the participation of PI3K in the FPRL1-mediated JNK, ERK and p38 pathways in both U87 and FPRL1/CHO cells due to their insensitivities to LY294002 pretreatment (Fig. 4).
Mitogen-activated protein kinases (MAPKs) are not only pivotal mediators of signal transduction but they also regulate diverse biological processes ranging from survival, proliferation and differentiation to apoptosis. By using human U87 astrocytoma and transfected FPRL1/CHO cells, we have demonstrated that activation of FPRL1 with WKYMVM effectively phosphorylated JNK and ERK. Interestingly, p38 MAPK activation was only seen with FPRL1/CHO cells. The MAPK phosphorylations in response to WKYMVM were blocked by WRW⁴ (a selective FPRL1 antagonist), but not cyclosporine H (a well-known FPR antagonist). The key signaling intermediates in the MAPK pathways were also delineated. G_i/G_o proteins, Src family tyrosine kinases, but not phosphatidylinositol-3 kinase, protein kinase C and calmodulin-dependent kinase II, were required to transmit signals from FPRL1 toward JNK, ERK and p38 MAPK. Furthermore, phospholipase Cβ was distinctively involved in the regulation of JNK but not the other MAPKs. Importantly, WKYMVM-stimulated U87 cells triggered noticeable increases in glial fibrillary acidic protein (GFAP) and interleukin-1α (IL-1α), which are correlated with reactive astrocytosis. In contrast, GFAP expression was not altered following stimulation with N-formyl-methionyl-leucyl-phenylalanine. Moreover, inhibitions of G_i/G_o proteins and JNK completely abolished both GFAP and IL-1α upregulations by FPRL1, while blockade of the MEK/ERK cascade exclusively suppressed the GFAP production. Consistently, overexpression of MEK1 and constitutively active JNKK in U87 cells led to ERK and JNK activation, respectively, which was accompanied with markedly increased GFAP production. We have thus identified a possible linkage among FPRL1, MAPKs, astrocytic activation and the inflammatory response.
Novel chemoattractant peptides for human leukocytes
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At this point, it is not clear whether WKYMVM and the novel peptides described in this study share the same unknown receptor or not. On the cell signaling pathways involved in the activation of ERK, we have already demonstrated that WKYMVM stimulates ERK activation via PI3K- and MEK-dependent but not PKC-dependent mechanism [50]. We also observed that stimulation of differentiated HL60 cells with 1 μM of WKYMVM caused ERK activation via PI3K- and MEK-dependent but not PKC-dependent mechanism (data not shown).
Phospholipase A₂ plays a key role in phagocytic cell functions. By screening a synthetic hexapeptide combinatorial library, we identified 24 novel peptides based on their ability to stimulate arachidonic acid release associated with cytosolic phospholipase A₂ activity in differentiated HL60 cells. The identified peptides, that contain the consensus sequence (K/R/M)KYY(P/V/Y)M, also induce intracellular calcium release in a pertussis toxin-sensitive manner showing specific action on phagocytic leukocytes, but not on other cells. Functionally, the peptides stimulate superoxide generation and chemotactic migration in human neutrophils and monocytes. Four of the tested active peptides were ligands for formyl peptide receptor like 1. Among these, two peptides with the consensus sequence (R/M)KYYYM can induce intracellular calcium release in undifferentiated HL60 cells that do not express formyl peptide receptor like 1, indicating usage of other receptor(s). A study of intracellular signaling in differentiated HL60 cells induced by the peptides has revealed that four of the novel peptides can induce extracellular signal-regulated protein kinase activation via shared and distinct signaling pathways, based on their dependence of phospatidylinositol-3-kinase, protein kinase C, and MEK. These peptides provide previously unavailable tools for study of differential signaling in leukocytes.
UVA Induces Ser<sup>381</sup> Phosphorylation of p90 <sup>RSK</sup>/MAPKAP-K1 via ERK and JNK Pathways
2001, Journal of Biological Chemistry
Citation Excerpt :
This suggests that besides ERKs-dependent pathways, p90RSK may also be activated by ERKs-independent pathways. Recently, full activation of the NTD kinase of p90RSK was shown to require cooperation of three kinases: ERKs (58, 59), the CTD kinase of p90RSK (16), and PDK1 (3-phosphoinositide-dependent protein kinase-1, a newly identified downstream kinase of PI 3-kinase). Ser221 in the activation loop of the NTD of p90RSK is known to be phosphorylated by PDK1 and this phosphorylation is proven to be essential for the activation of all p90RSK isoforms (29,60-62).
UVA exposure plays an important role in the etiology of skin cancer. The family of p90-kDa ribosomal S6 kinases (p90^RSK/MAPKAP-K1) are activated via phosphorylation. In this study, results show that UVA-induced phosphorylation of p90^RSK at Ser³⁸¹ through ERKs and JNKs, but not p38 kinase pathways. We provide evidence that UVA-induced p90^RSK phosphorylation and kinase activity were time- and dose-dependent. Both PD98059 and a dominant negative mutant of ERK2 blocked ERKs and p90^RSK Ser³⁸¹ phosphorylation, as well as p90^RSK activity. A dominant negative mutant of p38 kinase blocked UVA-induced phosphorylation of p38 kinase, but had no effect on UVA-induced Ser³⁸¹ phosphorylation of p90^RSK or kinase activity. UVA-induced p90^RSK phosphorylation and kinase activity were markedly attenuated in JnK₁−/− andJnK₂−/− cells. A dominant negative mutant of JNK₁ inhibited UVA-induced JNKs and p90^RSK phosphorylation and kinase activity, but had no effect on ERKs phosphorylation. PD169316, a novel inhibitor of JNKs and p38 kinase, inhibited phosphorylation of p90^RSK, JNKs, and p38 kinase, but not ERKs. However, SB202190, a selective inhibitor of p38 kinase, had no effect on p90^RSK or JNKs phosphorylation. Significantly, ERKs and JNKs, but not p38 kinase, immunoprecipitated with p90^RSK when stimulated by UVA and p90^RSK was a substrate for ERK2 and JNK2, but not p38 kinase. These data indicate clearly that p90^RSKSer³⁸¹ may be phosphorylated by activation of JNKs or ERKs, but not p38 kinase.
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Trp-Lys-Tyr-Met-Val-Met activates mitogen-activated protein kinase via a PI-3 kinase-mediated pathway independent of PKC

Abstract

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Blood

J. Biol. Chem.

Cell

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Clin. Biochem.

Biochem. Pharmacol.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol Chem.

J. Biol Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.