Diacylglycerol Kinase-α Mediates Hepatocyte Growth Factor-induced Epithelial Cell Scatter by Regulating Rac Activation and Membrane Ruffling

Chianale, Federica; Cutrupi, Santina; Rainero, Elena; Baldanzi, Gianluca; Porporato, Paolo E.; Traini, Sara; Filigheddu, Nicoletta; Gnocchi, Viola F.; Santoro, Massimo; Parolini, Ornella; Blitterswijk, Wim J. van; Sinigaglia, Fabiola; Graziani, Andrea

doi:10.1091/mbc.e07-02-0177

Cited by 32 publications

(39 citation statements)

References 76 publications

(89 reference statements)

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“…6 and Fig. S9) and Rac activation (13). Collectively, the data presented above identify DGKα as an upstream regulator of aPKCζ/ι.…”

Section: Dgkα Regulates Rac By Directing Its Recruitment To the Plasmamentioning

confidence: 53%

“…Culturing, fixing, and staining detailed procedures are described in SI Materials and Methods and ref. 13. Representative pictures are shown in the figures.…”

Section: Methodsmentioning

confidence: 99%

“…At early time points from HGF stimulation, cells at the periphery of a colony reorganize the actin cytoskeleton and form dynamic protrusions of the outer plasma membrane known as ruffles, mediated by membrane targeting and activation of Rac (14,15). We previously showed that downregulation of DGKα affected both HGF-induced membrane ruffling and recruitment of Rac to the plasma membrane (13). Here, by live-cell imaging, we report that HGF-induced rapid and transient recruitment of EGFP-Rac at sites of intense rufAuthor contributions: F.C., E.R., F.S., G.B., and A.G. designed research; F.C., E.R., C.C., V.B., A.P., G.S., and G.B.…”

Section: Dgkα Regulates Rac By Directing Its Recruitment To the Plasmamentioning

confidence: 99%

“…Activation of DGKα mediates growth-factor-induced cell migration and proliferation in epithelial, endothelial, and lymphoma cells (8)(9)(10)(11)(12). Moreover, we demonstrated that in epithelial cells, DGKα is required for hepatocyte growth factor (HGF) -induced membrane ruffling by regulating Rac membrane targeting and activation (13). However, the molecular mechanisms by which DGKα regulates Rac function still remain to be elucidated.…”

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confidence: 99%

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Diacylglycerol kinase α mediates HGF-induced Rac activation and membrane ruffling by regulating atypical PKC and RhoGDI

Chianale

Rainero

Cianflone

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Diacylglycerol kinases (DGKs) convert diacylglycerol (DAG) into phosphatidic acid (PA), acting as molecular switches between DAG-and PA-mediated signaling. We previously showed that Srcdependent activation and plasma membrane recruitment of DGKα are required for growth-factor-induced cell migration and ruffling, through the control of Rac small-GTPase activation and plasma membrane localization. Herein we unveil a signaling pathway through which DGKα coordinates the localization of Rac. We show that upon hepatocyte growth-factor stimulation, DGKα, by producing PA, provides a key signal to recruit atypical PKCζ/ι (aPKCζ/ι) in complex with RhoGDI and Rac at ruffling sites of colony-growing epithelial cells. Then, DGKα-dependent activation of aPKCζ/ι mediates the release of Rac from the inhibitory complex with RhoGDI, allowing its activation and leading to formation of membrane ruffles, which constitute essential requirements for cell migration. These findings highlight DGKα as the central element of a lipid signaling pathway linking tyrosine kinase growth-factor receptors to regulation of aPKCs and RhoGDI, and providing a positional signal regulating Rac association to the plasma membrane.cell migration | growth factors | phosphatidic acid C ell migration, central to many biological and pathological processes such as cancer metastatic progression, is a multistep cycle involving extension of protrusions and formation of stable attachments near the leading edge, followed by translocation of the cell body forward (1). The protrusive activity occurring at the leading edge depends on the spatial and temporal coordination between cell substrate adhesion and actin reorganization. Rhofamily small GTPases coordinate the recruitment at the leading edge of downstream effectors, thereby mediating the formation of ruffles and lamellipodia. Their GTP-bound state is tightly regulated by both guanine nucleotide exchange factors (GEFs), which stimulate GTP loading, and GTPase activating proteins (GAPs), which catalyze GTP hydrolysis. Moreover, Rho-family GTPases are regulated by guanine nucleotide dissociation inhibitors (GDIs), which antagonize both GEFs and GAPs and mediate the cycling of Rho proteins between the cytosol and the membrane (2, 3).Atypical protein kinase C ζ and ι (aPKCζ/ι), unlike classical and novel PKCs, feature a C1-like domain which does not bind to either diacylglycerol or phorbol esters, and have recently been proposed as key transducers for establishment of cell polarity and migration (4).Diacylglycerol kinases (DGKs), which convert diacylglycerol (DAG) to phosphatidic acid (PA), comprise a family of 10 distinct enzymes grouped into five classes, each featuring distinct regulatory domains and a highly conserved catalytic domain preceded by two cysteine-rich C1-like domains. An increasing body of evidence indicates that DGKs, by acting as terminators of diacylglyceroltriggered signaling, contribute to regulating C1 domain-containing proteins, such as classical and novel PKCs and the Rac-GAP β-chimaerin (5). ...

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“…6 and Fig. S9) and Rac activation (13). Collectively, the data presented above identify DGKα as an upstream regulator of aPKCζ/ι.…”

Section: Dgkα Regulates Rac By Directing Its Recruitment To the Plasmamentioning

confidence: 53%

“…Culturing, fixing, and staining detailed procedures are described in SI Materials and Methods and ref. 13. Representative pictures are shown in the figures.…”

Section: Methodsmentioning

confidence: 99%

Section: Dgkα Regulates Rac By Directing Its Recruitment To the Plasmamentioning

confidence: 99%

mentioning

confidence: 99%

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Diacylglycerol kinase α mediates HGF-induced Rac activation and membrane ruffling by regulating atypical PKC and RhoGDI

Chianale

Rainero

Cianflone

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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“…DGK activation has been reported to be required for hepatocyte growth factor-and v-Src-induced scattering and migration of epithelial cells ( 49 ). It is likely that signaling from cell-cell adhesion downregulates DGK activity and leads to a decline in PA concentration, which may impair cell scatter and migration.…”

Section: Pa Content In Hek293 Cellsmentioning

confidence: 99%