Abstract
PTEN is a tumor suppressor with dual protein and lipid–phosphatase activity, which is frequently deleted or mutated in many human advanced cancers. Recent studies have also demonstrated that PTEN is a promising target in type II diabetes and obesity treatment. Using C-terminal PTEN sequence in pEG202–NLS as bait, yeast two-hybrid screening on Mouse Embryo, Colon Cancer, and HeLa cDNA libraries was carried out. Isolated positive clones were validated by mating assay and identified through automated DNA sequencing and BLAST database searches. Sequence analysis revealed a number of PTEN-binding proteins linking this phosphatase to a number of different signaling cascades, suggesting that PTEN may perform other functions besides tumor-suppressing activity in different cell types. In particular, the interplay between PTEN function and adipocyte-specific fatty-acid-binding protein FABP4 is of notable interest. The demonstrable tautology of PTEN to FABP4 suggested a role for this phosphatase in the regulation of lipid metabolism and adipocyte differentiation. This interaction was further studied using coimmunoprecipitation and gel-filtration assays. Finally, based on Biacore assay, we have calculated the K D of PTEN–FABP4 complex, which is around 2.8 μM.
References
Simpson L, Parsons R (2001) PTEN: life as a tumor suppressor. Exp Cell Res 264:29–41. doi:10.1006/excr.2000.5130
Sulis ML, Parsons R (2003) PTEN: from pathology to biology. Trends Cell Biol 13:478–483. doi:10.1016/S0962-8924(03)00175-2
Tamguney T, Stokoe DJ (2007) New insights into PTEN. Cell Sci 120:4071–4079. doi:10.1242/10.1242/jcs.015230
Myers MP, Stolarov JP, Eng C, Li J, Wang SI, Wigler MH, Parsons R, Tonks NK (1997) P-TEN, the tumor suppressor from human chromosome 10q23, is a dual-specificity phosphatase. Proc Natl Acad Sci USA 94:9052–9057
Gericke A, Munson M, Ross AH (2006) Regulation of the PTEN phosphatase. Gene 374:1–9. doi:10.1016/j.jene.2006.02.024
Maehama T, Dixon JE (1998) The tumor suppressor, PTEN/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3, 4, 5-trisphosphate. J Biol Chem 273:13375–13378. doi:10.1074/jbc.273.22.13375
Gu J, Tamura M, Pankov R, Danen EH, Takino T, Matsumoto K, Yamada KM (1999) Shc and FAK differentially regulate cell motility and directionality modulated by PTEN. J Cell Biol 146:389–403
Mahimainathan L, Choudhury GG (2004) Inactivation of platelet-derived growth factor receptor by the tumor suppressor PTEN provides a novel mechanism of action of the phosphatase. J Biol Chem 279:15258–15268. doi:10.1074/jbc.M314328200
Wang X, Jiang X (2008) Post-translational regulation of PTEN. Oncogene 27:5454–5463. doi:10.1038/onc.2008.242
Okamura H, Yoshida K, Morimoto H, Haneji T (2005) PTEN expression elicited by EGR-1 transcription factor in calyculin A-induced apoptotic cells. J Cell Biochem 94:117–125. doi:10.1002/jcb.20283
Patel L, Pass I, Coxon P, Downes CP, Smith SA, Macphee CH (2001) Tumor suppressor and anti-inflammatory actions of PPARgamma agonists are mediated via upregulation of PTEN. Curr Biol 11:764–768. doi:10.1016/S0960-9822(01)00225-1
Teresi RE, Shaiu C-W, Chen C-S, Chatterjee VK, Waite KA, Eng C (2006) Increased PTEN expression due to transcriptional activation of PPARγ by lovastatin and rosiglitazone. Int J Cancer 118:2390–2398. doi:10.1002/ijc.21799
Becker S, Dossus L, Kaaks R (2009) Obesity related hyperinsulinaemia and hyperglycaemia and cancer development. Arch Physiol Biochem 115:86–96. doi:10.1080/13813450902878054
Lazar DF, Saltiel AR (2006) Lipid phosphatases as drug discovery targets for type 2 diabetes. Nat Rev Drug Discov 5:333–342. doi:10.1038/nrd2007
Kim SK, Novak RF (2007) The role of intracellular signaling in insulin-mediated regulation of drug metabolizing enzyme gene and protein expression. Pharmacol Ther 113:88–120. doi:10.1016/j.pharmthera.2006.07.004
Stiles B, Wang Y, Stahl A, Bassilian S, Lee WP, Kim YJ, Sherwin R, Devaskar S, Lesche R, Magnuson MA, Wu H (2004) Liver-specific deletion of negative regulator Pten results in fatty liver and insulin hypersensitivity. Proc Natl Acad Sci USA 101:2082–2087. doi:10.1073/pnas.0308617100
Wijesekara N, Konrad D, Eweida M, Jefferies C, Liadis N, Giacca A, Crackower M, Suzuki A, Mak TW, Kahn CR, Klip A, Woo M (2005) Muscle-specific Pten deletion protects against insulin resistance and diabetes. Mol Cell Biol 25:1135–1145. doi:10.1128/MCB.25.3.1135-1145.2005
Kurlawalla-Martinez C, Stiles B, Wang Y, Devaskar SU, Kahn BB, Wu H (2005) Insulin hypersensitivity and resistance to streptozotocin-induced diabetes in mice lacking PTEN in adipose tissue. Mol Cell Biol 25:2498–2510. doi:10.1128/MCB.25.6.2498-2510.2005
Gorbenko O, Kuznetsov V, Kukharenko O, Zhyvoloup A, Panasyuk G, Nemazanyy I, Filonenko V, Gout I (2004) Identification of a novel binding partners for tumor suppressor PTEN by a yeast two-hybrid approach. Eksp Onkol 26:15–19
Ro HS, Zhang L, Majdalawieh A, Kim SW, Wu X, Lyons PJ, Webber C, Ma H, Reidy SP, Boudreau A, Miller JR, Mitchell P, McLeod RS (2007) Adipocyte enhancer-binding protein 1 modulates adiposity and energy homeostasis. Obesity 15:288–302. doi:10.1038/oby.2007.569
Panasyuk G, Nemazanyy I, Filonenko V, Zhyvoloup A (2004) Large-scale yeast transformation in low-percentage agarose medium. Biotechniques 36:40–44
Gorbenko O, Filonenko V, Gout I (2006) Generation and characterization of monoclonal antibodies against FABP4. Hybridoma 25:86–90. doi:10.1089/hyb.2006.25.86
Jonsson U, Fagerstam L, Ivarsson B, Johnsson B, Karlsson R, Lundh K, Lofas S, Persson B, Roos H, Ronnberg I (1991) Real-time biospecific interaction analysis using surface plasmon resonance and a sensor chip technology. Biotechniques 11:620–627
Leslie NR, Gray A, Pass I, Orchiston EA, Downes CP (2000) Analysis of the cellular functions of PTEN using catalytic domain and C-terminal mutations: differential effects of C-terminal deletion on signalling pathways downstream of phosphoinositide 3-kinase. Biochem J 346:827–833
Okumura K, Zhao M, DePinho RA, Furnari FB, Cavenee WK (2005) Cellular transformation by the MSP58 oncogene is inhibited by its physical interaction with the PTEN tumor suppressor. Cell Cycle 4:540–542. doi:10.1073/pnas.0409370102
Fu Y, Luo N, Lopes-Virella MF, Garvey WT (2002) The adipocyte lipid binding protein (ALBP/aP2) gene facilitates foam cell formation in human THP-1 macrophages. Atherosclerosis 165:259–269
Makowski L, Hotamisligil GS (2005) The role of fatty acid binding proteins in metabolic syndrome and atherosclerosis. Curr Opin Lipidol 16:543–548
Uysal KT, Scheja L, Wiesbrock SM, Bonner-Weir S, Hotamisligil GS (2000) Improved glucose and lipid metabolism in genetically obese mice lacking aP2. Endocrinology 141:3388–3396
Furuhashi M, Tuncman G, Görgün CZ, Makowski L, Atsumi G, Vaillancourt E, Kono K, Babaev VR, Fazio S, Linton MF, Sulsky R, Robl JA, Parker RA, Hotamisligil GS (2007) Treatment of diabetes and atherosclerosis by inhibiting fatty-acid-binding protein aP2. Nature 447:959–965. doi:10.1038/nature05844
Tsuda M, Inoue-Narita T, Suzuki A, Itami S, Blumenberg M, Manabe M (2009) Induction of gene encoding FABP4 in Pten-null keratinocytes. FEBS Lett 583:1319–1322. doi:10.1016/j.febslet.2009.03.030
Horie Y, Suzuki A, Kataoka E, Sasaki T, Hamada K, Sasaki J, Mizuno K, Hasegawa G, Kishimoto H, Iizuka M, Naito M, Enomoto K, Watanabe S, Mak TW, Nakano T (2004) Hepatocyte-specific Pten deficiency results in steatohepatitis and hepatocellular carcinomas. J Clin Investig 113:1774–1783. doi:10.1172/JCI200420513
Boiteux G, Lascombe I, Roche E, Plissonnier ML, Clairotte A, Bittard H, Fauconnet S (2009) A-FABP, a candidate progression marker of human transitional cell carcinoma of the bladder, is differentially regulated by PPAR in urothelial cancer cells. Int J Cancer 15:1820–1828. doi:10.1002/ijc.24112
Hackl H, Maurer M, Mlecnik B, Hartler J, Stocker G, Miranda-Saavedra D, Trajanoski Z (2004) GOLD.db: genomics of lipid-associated disorders database. BMC Genomics 5:93. doi:10.1186/1471-2164-5-93
Vazquez F, Grossman SR, Takahashi Y, Rokas MV, Nakamura N, Sellers WR (2001) Phosphorylation of the PTEN tail acts as an inhibitory switch by preventing its recruitment into a protein complex. J Biol Chem 276:48627–48630. doi:10.1074/jbc.C100556200
Acknowledgment
This study was supported in part by Short-term Fellowships from FEBS, EMBO, and EACR. The authors thank Dr. Robert James for his careful reading of the manuscript.
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Gorbenko, O., Panayotou, G., Zhyvoloup, A. et al. Identification of novel PTEN-binding partners: PTEN interaction with fatty acid binding protein FABP4. Mol Cell Biochem 337, 299–305 (2010). https://doi.org/10.1007/s11010-009-0312-1
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DOI: https://doi.org/10.1007/s11010-009-0312-1