A signalling cascade involving receptor-activated phospholipase A

Animals Binding Sites Cell Movement Epidermal Growth Factor / pharmacology ErbB Receptors / metabolism Inositol Phosphates / metabolism Mice NIH 3T3 Cells Phospholipases A2 / metabolism Phosphorylation Protein Binding Protein Tyrosine Phosphatase, Non-Receptor Type 6 / chemistry RAW 264.7 Cells Signal Transduction Wound Healing src Homology Domains src-Family Kinases / metabolism
Actin polymerisation Cell motility EGF Glycerophosphoinositols Membrane ruffles Phosphoinositides SH2 domain Shp1

Journal

Cell communication and signaling : CCS
ISSN: 1478-811X
Titre abrégé: Cell Commun Signal
Pays: England
ID NLM: 101170464

Informations de publication

Date de publication:
01 03 2019
Historique:
received: 12 12 2018
accepted: 13 02 2019
entrez: 3 3 2019
pubmed: 3 3 2019
medline: 14 6 2019
Statut: epublish

Résumé

Shp1, a tyrosine-phosphatase-1 containing the Src-homology 2 (SH2) domain, is involved in inflammatory and immune reactions, where it regulates diverse signalling pathways, usually by limiting cell responses through dephosphorylation of target molecules. Moreover, Shp1 regulates actin dynamics. One Shp1 target is Src, which controls many cellular functions including actin dynamics. Src has been previously shown to be activated by a signalling cascade initiated by the cytosolic-phospholipase A Affinity chromatography, mass spectrometry and co-immunoprecipitation studies were employed to identify the GroPIns4P-interactors; among these Shp1 was selected for further analysis. The specific Shp1 residues interacting with GroPIns4P were revealed by NMR and validated by site-directed mutagenesis and biophysical methods such as circular dichroism, isothermal calorimetry, fluorescence spectroscopy, surface plasmon resonance and computational modelling. Morphological and motility assays were performed in NIH3T3 fibroblasts. We find that Shp1 is the direct cellular target of GroPIns4P. GroPIns4P directly binds to the Shp1-SH2 domain region (with the crucial residues being Ser 118, Arg 138 and Ser 140) and thereby promotes the association between Shp1 and Src, and the dephosphorylation of the Src-inhibitory phosphotyrosine in position 530, resulting in Src activation. As a consequence, fibroblast cells exposed to GroPIns4P show significantly enhanced wound healing capability, indicating that GroPIns4P has a stimulatory role to activate fibroblast migration. GroPIns4P is produced by cPLA This study identifies a so-far undescribed mechanism of Shp1/Src modulation that promotes cell motility and that is dependent on the cPLA

Sections du résumé

BACKGROUND
Shp1, a tyrosine-phosphatase-1 containing the Src-homology 2 (SH2) domain, is involved in inflammatory and immune reactions, where it regulates diverse signalling pathways, usually by limiting cell responses through dephosphorylation of target molecules. Moreover, Shp1 regulates actin dynamics. One Shp1 target is Src, which controls many cellular functions including actin dynamics. Src has been previously shown to be activated by a signalling cascade initiated by the cytosolic-phospholipase A
METHODS
Affinity chromatography, mass spectrometry and co-immunoprecipitation studies were employed to identify the GroPIns4P-interactors; among these Shp1 was selected for further analysis. The specific Shp1 residues interacting with GroPIns4P were revealed by NMR and validated by site-directed mutagenesis and biophysical methods such as circular dichroism, isothermal calorimetry, fluorescence spectroscopy, surface plasmon resonance and computational modelling. Morphological and motility assays were performed in NIH3T3 fibroblasts.
RESULTS
We find that Shp1 is the direct cellular target of GroPIns4P. GroPIns4P directly binds to the Shp1-SH2 domain region (with the crucial residues being Ser 118, Arg 138 and Ser 140) and thereby promotes the association between Shp1 and Src, and the dephosphorylation of the Src-inhibitory phosphotyrosine in position 530, resulting in Src activation. As a consequence, fibroblast cells exposed to GroPIns4P show significantly enhanced wound healing capability, indicating that GroPIns4P has a stimulatory role to activate fibroblast migration. GroPIns4P is produced by cPLA
CONCLUSIONS
This study identifies a so-far undescribed mechanism of Shp1/Src modulation that promotes cell motility and that is dependent on the cPLA

Identifiants

DOI: 10.1186/s12964-019-0329-3 PMID: 30823936 PMC: PMC6396489
pubmed: 30823936
doi: 10.1186/s12964-019-0329-3
pii: 10.1186/s12964-019-0329-3
pmc: PMC6396489
doi:

Substances chimiques

Inositol Phosphates 0
glycero-3-phosphoinositol 4-phosphate 125638-70-2
Epidermal Growth Factor 62229-50-9
ErbB Receptors EC 2.7.10.1
src-Family Kinases EC 2.7.10.2
Phospholipases A2 EC 3.1.1.4
Protein Tyrosine Phosphatase, Non-Receptor Type 6 EC 3.1.3.48
Ptpn6 protein, mouse EC 3.1.3.48

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

20

Subventions

Organisme : Associazione Italiana per la Ricerca sul Cancro
ID : IG10341 and IG18776
Pays : International
Organisme : Associazione Italiana per la Ricerca sul Cancro
ID : 20786
Pays : International

Références

Annu Rev Pharmacol Toxicol. 1999;39:175-89
pubmed: 10331081
J Immunol. 2000 Feb 1;164(3):1521-8
pubmed: 10640770
J Med Chem. 2000 Mar 23;43(6):1041-4
pubmed: 10737736
Semin Immunol. 2000 Aug;12(4):361-78
pubmed: 10995583
Mol Biol Cell. 2001 Jul;12(7):1937-56
pubmed: 11451994
Trends Cell Biol. 2002 Mar;12(3):112-20
pubmed: 11859023
Biochim Biophys Acta. 2002 May 23;1582(1-3):52-69
pubmed: 12069810
Eur J Biochem. 2002 Jun;269(12):3057-64
pubmed: 12071971
FASEB J. 2002 Nov;16(13):1823-5
pubmed: 12354693
J Biol Chem. 2003 Feb 14;278(7):4668-74
pubmed: 12468540
J Biol Chem. 2003 Feb 21;278(8):6516-20
pubmed: 12482860
J Am Chem Soc. 2003 Feb 19;125(7):1731-7
pubmed: 12580598
FEBS Lett. 2003 Feb 11;536(1-3):71-6
pubmed: 12586341
Mol Biol Cell. 2003 Feb;14(2):503-15
pubmed: 12589050
Exp Cell Res. 1958 Oct;15(2):332-45
pubmed: 13597891
Chem Rev. 2004 Aug;104(8):3607-22
pubmed: 15303830
Biochem Biophys Res Commun. 2005 May 27;331(1):1-14
pubmed: 15845350
Cell Signal. 1991;3(4):321-32
pubmed: 1657098
Chem Biol Drug Des. 2006 Mar;67(3):230-7
pubmed: 16611216
Mol Pharmacol. 2006 Aug;70(2):562-70
pubmed: 16717135
FASEB J. 2006 Dec;20(14):2567-9
pubmed: 17060404
FEBS Lett. 2006 Dec 22;580(30):6789-96
pubmed: 17141226
Cell Signal. 2007 Nov;19(11):2351-60
pubmed: 17716865
Cancer Res. 2007 Dec 15;67(24):11769-78
pubmed: 18089807
FEBS J. 2008 Mar;275(5):867-82
pubmed: 18298793
Biochim Biophys Acta. 2008 Dec;1783(12):2311-22
pubmed: 18722484
Mol Endocrinol. 2009 Jun;23(6):901-13
pubmed: 19264842
Cell Mol Life Sci. 2009 Nov;66(21):3449-67
pubmed: 19669618
J Pharmacol Sci. 2009 Oct;111(2):182-92
pubmed: 19783865
J Immunol. 2010 Jun 1;184(11):6529-36
pubmed: 20421638
Invest Ophthalmol Vis Sci. 2010 Nov;51(11):5601-9
pubmed: 20538990
Biochem Soc Trans. 2012 Feb;40(1):101-7
pubmed: 22260673
J Biol Chem. 2012 May 11;287(20):16849-59
pubmed: 22393044
Front Immunol. 2013 Jul 29;4:213
pubmed: 23908653
Cancer Sci. 2014 Mar;105(3):290-6
pubmed: 24350867
Int J Biochem Cell Biol. 2014 Jun;51:23-8
pubmed: 24681003
Nat Commun. 2014 Jun 30;5:4273
pubmed: 24978161
Cell Death Dis. 2015 May 07;6:e1750
pubmed: 25950480
Nat Rev Mol Cell Biol. 2015 Nov;16(11):691-8
pubmed: 26420231
Future Oncol. 2016 May;12(10):1287-98
pubmed: 26987952
Immunol Res. 2016 Aug;64(4):804-19
pubmed: 27216862
Int J Mol Sci. 2017 Jun 08;18(6):null
pubmed: 28594363
J Biol Chem. 2017 Aug 4;292(31):12828-12841
pubmed: 28600357
Curr Opin Cell Biol. 2018 Apr;51:97-102
pubmed: 29289897
Exp Cell Res. 1970 Jun;60(3):437-44
pubmed: 5463639
Eur J Biochem. 1995 Aug 1;231(3):593-601
pubmed: 7649158
Genomics. 1995 May 1;27(1):165-73
pubmed: 7665165
Cell. 1995 Jun 16;81(6):849-56
pubmed: 7781062
Cell. 1993 Jul 2;73(7):1445-54
pubmed: 8324828
J Biol Chem. 1993 Sep 25;268(27):20402-7
pubmed: 8397204
Cell. 1995 Dec 1;83(5):821-30
pubmed: 8521499
Biochem J. 1996 Jan 1;313 ( Pt 1):91-6
pubmed: 8546715
J Biol Chem. 1996 Apr 26;271(17):10385-90
pubmed: 8626611
Anal Biochem. 1996 May 1;236(2):275-83
pubmed: 8660505
Anticancer Res. 1996 May-Jun;16(3B):1341-50
pubmed: 8694500
Eur J Biochem. 1996 Oct 15;241(2):386-92
pubmed: 8917434
Biochemistry. 1997 Apr 15;36(15):4393-8
pubmed: 9109646
Mol Biol Cell. 1997 Mar;8(3):443-53
pubmed: 9188097
J Biol Chem. 1997 Aug 22;272(34):21113-9
pubmed: 9261115
Trends Biotechnol. 1997 Aug;15(8):302-7
pubmed: 9263478
J Biol Chem. 1997 Sep 12;272(37):23376-81
pubmed: 9287352
J Cell Sci. 1997 Oct;110 ( Pt 19):2449-59
pubmed: 9410883
Genetics. 1998 Aug;149(4):1707-15
pubmed: 9691030

Auteurs

Alessia Varone (A)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy. a.varone@ibp.cnr.it.

Stefania Mariggiò (S)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Manpreet Patheja (M)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Vincenzo Maione (V)

Magnetic Resonance Centre (CERM), University of Florence, 50019, Sesto Fiorentino, Italy.

Antonio Varriale (A)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.
Institute of Food Science, National Research Council, Via Roma 64, 83100, Avellino, Italy.

Mariangela Vessichelli (M)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Daniela Spano (D)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Fabio Formiggini (F)

Italian Institute of Technology, Centre for Advanced Biomaterials for Health Care at CRIB, Largo Barsanti e Matteucci 53, 80125, Naples, Italy.

Matteo Lo Monte (M)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Nadia Brancati (N)

Institute of High Performance Computing and Networking, National Research Council, Via P. Castellino 111, 80131, Naples, Italy.

Maria Frucci (M)

Institute of High Performance Computing and Networking, National Research Council, Via P. Castellino 111, 80131, Naples, Italy.

Pompea Del Vecchio (P)

Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126, Naples, Italy.

Sabato D'Auria (S)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.
Institute of Food Science, National Research Council, Via Roma 64, 83100, Avellino, Italy.

Angela Flagiello (A)

CEINGE Advanced Biotechnology, Via G. Salvatore 486, 80145, Naples, Italy.

Clara Iannuzzi (C)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.
Department of Biochemistry, Biophysics and General Pathology, Second University of Naples, Via L. de Crecchio 7, 80138, Naples, Italy.

Alberto Luini (A)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Piero Pucci (P)

Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126, Naples, Italy.
CEINGE Advanced Biotechnology, Via G. Salvatore 486, 80145, Naples, Italy.

Lucia Banci (L)

Magnetic Resonance Centre (CERM), University of Florence, 50019, Sesto Fiorentino, Italy.

Carmen Valente (C)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy.

Daniela Corda (D)

Institute of Protein Biochemistry, National Research Council, Via Pietro Castellino 111, 80131, Naples, Italy. d.corda@ibp.cnr.it.
ORCID: 0000-0002-3614-751X

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › A signalling cascade involving...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Sites de fixation › A signalling cascade involving...
questionsmedicales.fr › Phénomènes physiologiques › Mouvement cellulaire › A signalling cascade involving...
questionsmedicales.fr › Facteurs biologiques › Protéines et peptides de signalisation intercellulaire › Protéines de la famille de l'EGF › Facteur de croissance épidermique › A signalling cascade involving...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines membranaires › Récepteurs de surface cellulaire › Récepteurs peptidiques › Récepteur facteur croissance › Récepteurs ErbB › A signalling cascade involving...
questionsmedicales.fr › Glucides › Oses phosphates › Inositol phosphates › A signalling cascade involving...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › A signalling cascade involving...
questionsmedicales.fr › Cellules › Cellules du tissu conjonctif › Fibroblastes › Cellules 3T3 › Cellules NIH 3T3 › A signalling cascade involving...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Hydrolases › Esterases › Carboxylic ester hydrolases › Phospholipases A › Phospholipases A2 › A signalling cascade involving...
questionsmedicales.fr › Métabolisme › Phosphorylation › A signalling cascade involving...
questionsmedicales.fr › Métabolisme › Liaison aux protéines › A signalling cascade involving...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines et peptides de signalisation intracellulaire › Protein Tyrosine Phosphatases › SH2 Domain-Containing Protein Tyrosine Phosphatases › Protein Tyrosine Phosphatase, Non-Receptor Type 6 › A signalling cascade involving...
questionsmedicales.fr › Cellules › Cellules RAW 264.7 › A signalling cascade involving...
questionsmedicales.fr › Phénomènes physiologiques cellulaires › Transduction du signal › A signalling cascade involving...
questionsmedicales.fr › Phénomènes biologiques › Régénération › Cicatrisation de plaie › A signalling cascade involving...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Conformation moléculaire › Conformation des protéines › Éléments structuraux des protéines › Motifs et domaines d'intéraction protéique › Domaine d'homologie SRC › A signalling cascade involving...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines et peptides de signalisation intracellulaire › Protein-tyrosine kinases › src-Family kinases › A signalling cascade involving...