PKCα-mediated phosphorylation of the diacylglycerol kinase ζ MARCKS domain switches cell migration modes by regulating interactions with Rac1 and RhoA.
Animals
Cell Movement
Diacylglycerol Kinase
/ physiology
Diglycerides
/ metabolism
Dystrophin-Associated Proteins
/ genetics
Fibroblasts
/ cytology
Mice
Mice, Knockout
Myristoylated Alanine-Rich C Kinase Substrate
/ genetics
Neuropeptides
/ genetics
Protein Domains
Protein Kinase C-alpha
/ pharmacology
rac1 GTP-Binding Protein
/ genetics
rhoA GTP-Binding Protein
/ genetics
PDZ domain
Ras homolog gene family
Ras-related C3 botulinum toxin substrate 1 (Rac1)
diacylglycerol
diacylglycerol kinase (DGK, DAGK)
member A (RhoA)
phosphatidic acid
protein kinase C (PKC)
scaffold protein
syntrophin
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
09
07
2020
revised:
26
02
2021
accepted:
03
03
2021
pubmed:
8
3
2021
medline:
21
8
2021
entrez:
7
3
2021
Statut:
ppublish
Résumé
Cells can switch between Rac1 (lamellipodia-based) and RhoA (blebbing-based) migration modes, but the molecular mechanisms regulating this shift are not fully understood. Diacylglycerol kinase ζ (DGKζ), which phosphorylates diacylglycerol to yield phosphatidic acid, forms independent complexes with Rac1 and RhoA, selectively dissociating each from their common inhibitor RhoGDI. DGKζ catalytic activity is required for Rac1 dissociation but is dispensable for RhoA dissociation; instead, DGKζ stimulates RhoA release via a kinase-independent scaffolding mechanism. The molecular determinants that mediate the selective targeting of DGKζ to Rac1 or RhoA signaling complexes are unknown. Here, we show that protein kinase Cα (PKCα)-mediated phosphorylation of the DGKζ MARCKS domain increased DGKζ association with RhoA and decreased its interaction with Rac1. The same modification also enhanced DGKζ interaction with the scaffold protein syntrophin. Expression of a phosphomimetic DGKζ mutant stimulated membrane blebbing in mouse embryonic fibroblasts and C2C12 myoblasts, which was augmented by inhibition of endogenous Rac1. DGKζ expression in differentiated C2 myotubes, which have low endogenous Rac1 levels, also induced substantial membrane blebbing via the RhoA-ROCK pathway. These events were independent of DGKζ catalytic activity, but dependent upon a functional C-terminal PDZ-binding motif. Rescue of RhoA activity in DGKζ-null cells also required the PDZ-binding motif, suggesting that syntrophin interaction is necessary for optimal RhoA activation. Collectively, our results define a switch-like mechanism whereby DGKζ phosphorylation by PKCα plays a role in the interconversion between Rac1 and RhoA signaling pathways that underlie different cellular migration modes.
Identifiants
pubmed: 33676892
pii: S0021-9258(21)00292-1
doi: 10.1016/j.jbc.2021.100516
pmc: PMC8042443
pii:
doi:
Substances chimiques
Diglycerides
0
Dystrophin-Associated Proteins
0
Marcks protein, mouse
0
Neuropeptides
0
Rac1 protein, mouse
0
syntrophin
0
Myristoylated Alanine-Rich C Kinase Substrate
125267-21-2
Diacylglycerol Kinase
EC 2.7.1.107
diacylglycerol kinase zeta, mouse
EC 2.7.1.107
Protein Kinase C-alpha
EC 2.7.11.13
RhoA protein, mouse
EC 3.6.5.2
rac1 GTP-Binding Protein
EC 3.6.5.2
rhoA GTP-Binding Protein
EC 3.6.5.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
100516Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
Références
Mol Biol Cell. 2009 Apr;20(7):2049-59
pubmed: 19211846
J Cell Sci. 2006 Apr 15;119(Pt 8):1528-36
pubmed: 16551696
PLoS One. 2015 Dec 23;10(12):e0144942
pubmed: 26701304
Mol Cell. 2004 Jul 2;15(1):117-27
pubmed: 15225553
Curr Biol. 1997 Aug 1;7(8):611-4
pubmed: 9259558
J Cell Biol. 2003 Mar 17;160(6):929-37
pubmed: 12629049
Nature. 1998 Aug 13;394(6694):697-700
pubmed: 9716136
EMBO J. 1999 Feb 1;18(3):578-85
pubmed: 9927417
Trends Cell Biol. 2011 Dec;21(12):718-26
pubmed: 21924908
J Biol Chem. 2010 Jul 23;285(30):23296-308
pubmed: 20472934
Annu Rev Biochem. 2003;72:743-81
pubmed: 12676796
Curr Opin Cell Biol. 2010 Oct;22(5):690-6
pubmed: 20829016
Small GTPases. 2012 Oct-Dec;3(4):219-24
pubmed: 22906997
Nature. 2002 Dec 12;420(6916):629-35
pubmed: 12478284
Mol Biol Cell. 2003 Nov;14(11):4499-511
pubmed: 14551255
J Biol Chem. 2002 Aug 16;277(33):30300-9
pubmed: 12015310
J Biol Chem. 2003 Oct 10;278(41):39542-7
pubmed: 12890670
FEBS Lett. 2008 Jun 18;582(14):2093-101
pubmed: 18460342
Mol Cell Biol. 2005 Aug;25(16):7289-302
pubmed: 16055737
Biochim Biophys Acta. 2009 Sep;1791(9):942-8
pubmed: 19264149
Cell. 1992 Aug 7;70(3):401-10
pubmed: 1643658
Small GTPases. 2018 Jul 4;9(4):316-321
pubmed: 27533896
Soc Gen Physiol Ser. 1997;52:197-207
pubmed: 9210230
Biochem Soc Trans. 1995 Aug;23(3):456-9
pubmed: 8566347
J Cell Sci. 1998 Oct;111 ( Pt 19):2911-22
pubmed: 9730983
Nature. 1993 Dec 16;366(6456):643-54
pubmed: 8259209
Trends Cell Biol. 2005 Jul;15(7):356-63
pubmed: 15921909
J Biol Chem. 2001 Oct 5;276(40):37307-16
pubmed: 11489882
Cell Syst. 2016 Jan 27;2(1):38-48
pubmed: 27136688
Mol Biol Cell. 2012 Oct;23(20):4008-19
pubmed: 22918940
Biochem J. 2005 Aug 15;390(Pt 1):1-9
pubmed: 16083425
Trends Biochem Sci. 1995 Jul;20(7):272-6
pubmed: 7667880
Cell. 2008 Oct 31;135(3):510-23
pubmed: 18984162
Cell. 1992 Nov 27;71(5):713-6
pubmed: 1423627
J Biol Chem. 2001 Jul 13;276(28):26526-33
pubmed: 11352924