Diacylglycerol kinase-α phosphorylation by Src on Y335 is required for activation, membrane recruitment and Hgf-induced cell motility

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

doi:10.1038/sj.onc.1210717

Cited by 50 publications

(49 citation statements)

References 40 publications

(69 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PKA and PKC have been shown to phosphorylate DGK in COS7 cells ( 85 ). Phosphorylation of DGK ␣ by the tyrosine kinase Src confers hepatocyte growth factor-induced cell motility ( 86,87 ), whereas PKC-catalyzed phosphorylation of DGK promotes the dissociation of the lipid kinase from PKC ( 88,89 ). Given that DAG stimulates PKC activity, the association with DGK provides a mechanism to limit the ability of PKC to phosphorylate target proteins.…”

Section: Discussionmentioning

confidence: 99%

cAMP-stimulated transcription of DGKθ requires steroidogenic factor 1 and sterol regulatory element binding protein 1

Cai

Sewer

2013

Journal of Lipid Research

View full text Add to dashboard Cite

( 4 ). DGK -null mice exhibit several neural abnormalities, including a higher resistance of electroconvulsive shock ( 5 ) and increased cyclooxygenase 2 and tyrosine hydroxylase expression ( 6 ), suggesting a role for DGK in regulating synaptic activity. Mice lacking DGK ␣ ( 7 ) or DGK ( 8 ) exhibit enhanced T cell function and demonstrate a role for these kinases in controlling DAG metabolism during the immune response. DGK isoforms have been implicated in various other cellular processes including inhibition of Rap1 signaling ( 9 ) and retinoblastoma-mediated cell cycle control ( 10 ). DGK is activated by nerve growth factor in PC12 cells ( 11 ) and thrombin in IIC9 fi broblasts ( 12, 13 ), whereas DGK promotes myogenesis in C2C12 cells ( 14 ) and DGK ␥ plays a role in regulating the cell cycle in CHO-K cells ( 15 ).To date, 10 mammalian DGKs have been identifi ed that are divided into fi ve groups based on functional domains ( 16,17 ). However, all isoforms contain cysteine-rich zinc fi nger-like structures, a conserved catalytic region ( 18-21 ). DGK , the sole member of group V, is comprised of three cysteine-rich domains (CRDs), a proline/glycine-rich domain at its N terminus, and a pleckstrin homology (PH) with an overlapping Ras-binding domain ( 22 ). While the functions of many of the other domains in DGK are unclear, the catalytic activity requires all domains of the enzyme ( 23 ). It has been postulated that the CRDs of the enzyme are required both for correct folding of the protein and for substrate presentation ( 23 ). Mutation of the CRD of DGK diminishes DAG-induced translocation of the enzyme to the plasma membrane ( 24 ); whereas the interaction between DGK and the nuclear receptor steroidogenic factor 1 (SF1) requires the PH domain ( 25 ). Abstract Diacylglycerol kinase (DGK) is

show abstract

Section: Discussionmentioning

confidence: 99%

cAMP-stimulated transcription of DGKθ requires steroidogenic factor 1 and sterol regulatory element binding protein 1

Cai

Sewer

2013

Journal of Lipid Research

View full text Add to dashboard Cite

show abstract

“…Experiments with overexpressed enzyme suggest that DGK␣ Y335 phosphorylation targets DGK␣ to the plasma membrane (37,38). Analysis of endogenous protein in Jurkat T lymphocytes demonstrates that tyrosine-phosphorylated DGK␣ represents a membrane-associated fraction, which probably corresponds to the active pool of the enzyme.…”

Section: Discussionmentioning

confidence: 99%

“…The physiological significance of this rapid loss of association remains to be determined, but it may favor DGK␣ association with other partners. A recent publication showed that phosphorylated Y335 might interact with certain SH2 domains in vitro (37). The rapid dissociation of DGK␣ pY335 and Lck might also control and/or restrict an inhibitory DGK␣ effect on Lck activity.…”

Section: Discussionmentioning

confidence: 99%

“…HEK293 cells do not express Lck, although they express other Src kinases that might phosphorylate the ectopically expressed DGK␣. Accordingly, Src kinases are reported to be essential for DGK␣ modulation in various adherent cell lines in response to receptors such as vascular endothelial growth factor, hepatocyte growth factor, or ␣-D-tocopherol (30,37,38,44,45).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Lck-Dependent Tyrosine Phosphorylation of Diacylglycerol Kinase α Regulates Its Membrane Association in T Cells

Merino

Ávila-Flores

Shirai³

et al. 2008

The Journal of Immunology

View full text Add to dashboard Cite

TCR engagement triggers phospholipase Cγ1 activation through the Lck-ZAP70-linker of activated T cell adaptor protein pathway. This leads to generation of diacylglycerol (DAG) and mobilization of intracellular Ca2+, both essential for TCR-dependent transcriptional responses. TCR ligation also elicits transient recruitment of DAG kinase α (DGKα) to the lymphocyte plasma membrane to phosphorylate DAG, facilitating termination of DAG-regulated signals. The precise mechanisms governing dynamic recruitment of DGKα to the membrane have not been fully elucidated, although Ca2+ influx and tyrosine kinase activation were proposed to be required. We show that DGKα is tyrosine phosphorylated, and identify tyrosine 335 (Y335), at the hinge between the atypical C1 domains and the catalytic region, as essential for membrane localization. Generation of an Ab that recognizes phosphorylated Y335 demonstrates Lck-dependent phosphorylation of endogenous DGKα during TCR activation and shows that pY335DGKα is a minor pool located exclusively at the plasma membrane. Our results identify Y335 as a residue critical for DGKα function and suggest a mechanism by which Lck-dependent phosphorylation and Ca2+ elevation regulate DGKα membrane localization. The concerted action of these two signals results in transient, receptor-regulated DGKα relocalization to the site at which it exerts its function as a negative modulator of DAG-dependent signals.

show abstract

“…We previously showed that DGKα is activated by growth factors on recruitment to the plasma membrane through its Src-mediated phosphorylation on Tyr 335 . 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).…”

mentioning

confidence: 99%

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.

View full text Add to dashboard Cite

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). ...

show abstract

Diacylglycerol kinase-α phosphorylation by Src on Y335 is required for activation, membrane recruitment and Hgf-induced cell motility

Cited by 50 publications

References 40 publications

cAMP-stimulated transcription of DGKθ requires steroidogenic factor 1 and sterol regulatory element binding protein 1

cAMP-stimulated transcription of DGKθ requires steroidogenic factor 1 and sterol regulatory element binding protein 1

Lck-Dependent Tyrosine Phosphorylation of Diacylglycerol Kinase α Regulates Its Membrane Association in T Cells

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

Contact Info

Product

Resources

About