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Filamin A Mutant Lacking Actin-Binding Domain Restores Mu Opioid Receptor Regulation in Melanoma Cells

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Abstract

We have previously reported that the protein filamin A (FLA) binds to the carboxyl tail of the mu opioid receptor (MOPr). Using human melanoma cells, which do not express filamin A, we showed that receptor down-regulation, functional desensitization and trafficking are deficient in the absence of FLA (Onoprishvili et al. Mol Pharmacol 64:1092–1100, 2003). Since FLA has a binding domain for actin and is a member of the family of actin cytoskeleton proteins, it is usually assumed that FLA functions via the actin cytoskeleton. We decided to test this hypothesis by preparing cDNA coding for mutant FLA lacking the actin binding domain (FLA-ABD) and expressing FLA-ABD in the human melanoma cell line M2 (M2-ABD cell line). We report here that this mutant is capable of restoring almost as well as full length FLA the down-regulation of the human MOPr. It is similarly very effective in restoring functional desensitization of MOPr, as assessed by the decrease in G-protein activation after chronic exposure of M2-ABD cells to the mu agonist DAMGO. We also found that A7 cells, expressing wild type FLA, exhibit rapid activation of the MAP kinases, ERK 1 and 2, by DAMGO, as shown by a rise in the level of phospho-ERK 1 and 2. This is followed by rapid dephosphorylation (inactivation), which reaches basal level between 30 and 60 min after DAMGO treatment. M2 cells show normal activation of ERK 1 and 2 in the presence of DAMGO, but very slow inactivation. The rapid rate of MAPK inactivation is partially restored by FLA-ABD. We conclude that some functions of FLA do not act via the actin cytoskeleton. It is likely that other functions, not studied here, may require functional binding of the MOPr-FLA complex to actin.

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Abbreviations

GPCR:

G protein-coupled receptor

FLA:

Filamin A

MOPr:

Mu opioid receptor

ABD:

Actin-binding domain

FLA-ABD:

FLA deletion mutant lacking ABD

DAMGO:

[d-Ala2, N-Me-Phe4,Gly5-ol]enkephalin

MAPK:

Mitogen-activated protein kinase

ERK:

Extracellular signal-regulated kinase (member of MAPK family)

GTPγS:

Guanosine 5′-3-O-(thio)triphosphate

M2-ABD:

M2 cells containing FLA-ABD cells

References

  1. Onoprishvili I, Andria ML, Kramer HK et al (2003) Interaction between the mu opioid receptor and filamin A is involved in receptor regulation and trafficking. Mol Pharmacol 64:1092–1100

    Article  PubMed  CAS  Google Scholar 

  2. Gorlin JB, Yamin R, Egan S et al (1990) Human endothelial actin-binding protein (ABP-280, nonmuscule filamin): a molecular leaf spring. J Cell Biol 111:1089–1105

    Article  PubMed  CAS  Google Scholar 

  3. Stossel TP, Condeelis J, Cooley L et al (2001) Filamins as integrators of cell mechanics and signalling. Nat Rev Mol Cell Biol 2:138–145

    Article  PubMed  CAS  Google Scholar 

  4. Loo DT, Kanner SB, Aruffo A (1998) Filamin binds to the cytoplasmic domain of the beta1-integrin. J Biol Chem 273:233004–223312

    Google Scholar 

  5. Xu BW, Xie ZW, Chung DW et al (1998) A novel human actin-binding protein homologue that binds to platelet glycoprotein Ibalpha. Blood 92:1268–1276

    PubMed  CAS  Google Scholar 

  6. Zhang J, Ferguson SSG, Barak LS et al (1998) Role for G protein-coupled receptor kinase in agonist-specific regulation of μ-opioid receptor responsiveness. Proc Natl Acad Sci USA 95:7157–7162

    Article  PubMed  CAS  Google Scholar 

  7. Onoprishvili I, Simon EJ (2007) Chronic morphine treatment up-regulates mu opioid receptor binding in cells lacking filamin A. Brain Res 1177:9–18

    Article  PubMed  CAS  Google Scholar 

  8. Gutkind JS (1998) The pathway connecting G-protein-coupled receptors to the nucleus through divergent mitogen-activated protein kinase cascades. J Biol Chem 273:1839–1842

    Article  PubMed  CAS  Google Scholar 

  9. Polakiewicz RD, Schieferl SM, Dorner LF et al (1998) A mitogen-activated protein kinase pathway is required for mu-opioid receptor desensitization. J Biol Chem 273:12402–12406

    Article  PubMed  CAS  Google Scholar 

  10. Trapaidze N, Gomes I, Cvejic S et al (2000) Opioid receptor endocytosis and activation of MAP kinase pathway. Brain Res Mol Brain Res 76:220–228

    Article  PubMed  CAS  Google Scholar 

  11. Kramer HK, Simon EJ (2000) Mu and delta opioid receptor agonists induce mitogen-activated protein kinase (MAPK): activation in the absence of internalization. Neuropharmacology 39:1707–1719

    Article  PubMed  CAS  Google Scholar 

  12. Schmidt H, Schulz S, Klutzny M et al (2000) Involvement of mitogen-activated protein kinase in agonist-induced phosphorylation of the mu opioid receptor in HEK 293 cells. J Neurochem 74:414–422

    Article  PubMed  CAS  Google Scholar 

  13. Kim EY, Ridgway LD, Dryer SE (2007) Interactions with filamin A stimulate surface expression of large-conductance Ca2+-activated K+ channels in the absence of direct actin binding. Mol Pharmacol 72:622–630

    Article  PubMed  CAS  Google Scholar 

  14. Sasaki A, Masuda Y, Ohta Y et al (2001) Filamin associates with Smads and regulates transforming growth factor-beta signaling. J Biol Chem 276:17871–17877

    Article  PubMed  CAS  Google Scholar 

  15. Li M, Bermak JC, Wang ZW et al (2000) Modulation of dopamine D2 receptor signaling by actin-binding protein (ABP-280). Mol Pharmacol 57:446–452

    PubMed  CAS  Google Scholar 

  16. Lin R, Karpa K, Kabbani N et al (2001) Dopamine D2 and D3 receptors are linked to the actin cytoskeleton via interaction with filamin A. Proc Natl Acad Sci USA 98:5258–5263

    Article  PubMed  CAS  Google Scholar 

  17. Cho EY, Cho DI, Park JH et al (2007) Roles of protein kinase C and actin-binding protein 280 in the regulation of intracellular trafficking of dopamine D3 receptor. Mol Endocrinol 21:2242–2254

    Article  PubMed  CAS  Google Scholar 

  18. Li M, Li C, Weingarten P et al (2002) Association of dopamine D3 receptors with actin-binding protein 280 (ABP-280). Biochem Pharmacol 63:859–863

    Article  PubMed  CAS  Google Scholar 

  19. Kim KM, Gainetdinov RR, Laporte SA et al (2005) G protein-coupled receptor kinase regulates dopamine D3 receptor signaling by modulating the stability of a receptor/filamin/beta-arrestin complex: a case of autoreceptor regulation. J Biol Chem (in press)

  20. Scott MG, Pierotti V, Storez H et al (2006) Cooperative regulation of extracellular signal-regulated kinase activation and cell shape change by filamin A and beta-arrestins. Mol Cell Biol 26:3432–3445

    Article  PubMed  CAS  Google Scholar 

  21. Hjalm G, MacLeod RJ, Kifor O et al (2001) Filamin-A binds to the carboxyl-terminal tail of the calcium-sensing receptor, an interaction that participates in CaR-mediated activation of mitogen-activated protein kinase. J Biol Chem 276:34880–34887

    Article  PubMed  CAS  Google Scholar 

  22. Awata H, Huang C, Handlogten ME et al (2001) Interaction of the calcium-sensing receptor and filamin, a potential scaffolding protein. J Biol Chem 276:34871–34879

    Article  PubMed  CAS  Google Scholar 

  23. Zhang M, Breitwieser GE (2005) High affinity interaction with filamin A protects against calcium-sensing receptor degradation. J Biol Chem 280:11140–11146

    Article  PubMed  CAS  Google Scholar 

  24. Huang C, Wu Z, Hujer KM et al (2006) Silencing of filamin A gene expression inhibits Ca2+-sensing receptor signaling. FEBS Lett 580:1795–1800

    Article  PubMed  CAS  Google Scholar 

  25. Seck T, Baron R, Horne WC (2003) Binding of filamin to the C-terminal tail of the calcitonin receptor controls recycling. J Biol Chem 278:10408–10416

    Article  PubMed  CAS  Google Scholar 

  26. He HJ, Kole S, Kwon YK et al (2003) Interaction of filamin A with the insulin receptor alters insulin-dependent activation of the mitogen-activated protein kinase pathway. J Biol Chem 278:27096–27104

    Article  PubMed  CAS  Google Scholar 

  27. Enz R (2002) The actin-binding protein filamin-A interatcs with the metabotropic glutamate receptor type 7. FEBS Lett 514:184–188

    Article  PubMed  CAS  Google Scholar 

  28. Enz R, Croci C (2003) Different binding motifs in metabotropic glutamate receptor type 7b for filamin A, protein phosphatase 1C, protein interacting with protein kinase C (PICK) 1 and syntenin allow the formation of multimeric protein complexes. Biochem J 372:183–191

    Article  PubMed  CAS  Google Scholar 

  29. Zhang T, Xu Q, Chen FR et al (2004) Yeast two-hybrid screening for proteins that interact with alpha1-adrenergic receptors. Acta Pharmacol Sin 25:1471–1478

    PubMed  CAS  Google Scholar 

  30. Nakamura F, Osborn TM, Hartemink CA et al (2007) Structural basis of filamin A functions. J Cell Biol 179:1011–1025

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Drs. Thomas P. Stossel, Y. Ohta and F. Nakamura for providing us with the M2 and A7 melanoma cells and the FLA cDNA construct. This work was supported by NIDA grant DA00017 to EJS and a Fellowship from NIDA Training Grant DA-07254 to IO.

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Authors and Affiliations

  1. Department of Psychiatry, New York University School of Medicine, 550 First Avenue, New York, NY, 10016, USA

    Irma Onoprishvili, Solav Ali, Matthew L. Andria, Adam Shpigel & Eric J. Simon

  2. Department of Pharmacology, New York University School of Medicine, 550 First Avenue, New York, NY, 10016, USA

    Eric J. Simon

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  1. Irma Onoprishvili
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  2. Solav Ali
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  3. Matthew L. Andria
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  4. Adam Shpigel
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  5. Eric J. Simon
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Correspondence to Eric J. Simon.

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Special issue article in honor of Dr. Ji-Sheng Han.

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Onoprishvili, I., Ali, S., Andria, M.L. et al. Filamin A Mutant Lacking Actin-Binding Domain Restores Mu Opioid Receptor Regulation in Melanoma Cells. Neurochem Res 33, 2054–2061 (2008). https://doi.org/10.1007/s11064-008-9684-y

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  • DOI: https://doi.org/10.1007/s11064-008-9684-y

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