Phospholipase A/Acyltransferase enzyme activity of H-rev107 inhibits the H-RAS signaling pathway

Journal of Biomedical Science

Fu-Ming Tsai¹  Shun-Yuan Jiang¹  Mao-Liang Chen¹  Lu-Kai Wang²  Tzung-Chieh Tsai³  Chang-Chieh Wu⁶  Rong-Yaun Shyu⁴  Chun-Hua Wang⁵ 

[1] Department of Research, Taipei Tzuchi Hospital, The Buddhist Tzuchi Medical Foundation, New Taipei City, Taiwan;Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan;Department of Microbiology, Immunology and Biopharmaceuticals, National Chiayi University, Chiayi, Taiwan;School of Medicine, Tzu Chi University, Hualien, Taiwan;Department of Dermatology, Taipei Tzuchi Hospital, The Buddhist Tzuchi Medical Foundation, New Taipei City, Taiwan;Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan

关键词: Acyl-biotin exchange assay;    Phospholipase A/acyltransferase;    H-RAS;    PLA2G16;    HRASLS3;    H-rev107;   

Others  :  817656
DOI  :  10.1186/1423-0127-21-36

 received in 2013-12-13, accepted in 2014-04-24,  发布年份 2014

【 摘 要 】

Background

H-rev107, also called HRASLS3 or PLA2G16, is a member of the HREV107 type II tumor suppressor gene family. Previous studies showed that H-rev107 exhibits phospholipase A/acyltransferase (PLA/AT) activity and downregulates H-RAS expression. However, the mode of action and the site of inhibition of H-RAS by H-rev107 are still unknown.

Results

Our results indicate that H-rev107 was co-precipitated with H-RAS and downregulated the levels of activated RAS (RAS-GTP) and ELK1-mediated transactivation in epidermal growth factor-stimulated and H-RAS-cotransfected HtTA cervical cancer cells. Furthermore, an acyl-biotin exchange assay demonstrated that H-rev107 reduced H-RAS palmitoylation. H-rev107 has been shown to be a PLA/AT that is involved in phospholipid metabolism. Treating cells with the PLA/AT inhibitor arachidonyl trifluoromethyl ketone (AACOCF3) or methyl arachidonyl fluorophosphate (MAFP) alleviated H-rev107-induced downregulation of the levels of acylated H-RAS. AACOCF3 and MAFP also increased activated RAS and ELK1-mediated transactivation in H-rev107-expressing HtTA cells following their treatment with epidermal growth factor. In contrast, treating cells with the acyl-protein thioesterase inhibitor palmostatin B enhanced H-rev107-mediated downregulation of acylated H-RAS in H-rev107-expressing cells. Palmostatin B had no effect on H-rev107-induced suppression of RAS-GTP levels or ELK1-mediated transactivation. These results suggest that H-rev107 decreases H-RAS activity through its PLA/AT activity to modulate H-RAS acylation.

Conclusions

We made the novel observation that H-rev107 decrease in the steady state levels of H-RAS palmitoylation through the phospholipase A/acyltransferase activity. H-rev107 is likely to suppress activation of the RAS signaling pathway by reducing the levels of palmitoylated H-RAS, which decreases the levels of GTP-bound H-RAS and also the activation of downstream molecules. Our study further suggests that the PLA/AT activity of H-rev107 may play an important role in H-rev107-mediated RAS suppression.

【 授权许可】

   
2014 Wang et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Husmann K, Sers C, Fietze E, Mincheva A, Lichter P, Schafer R: Transcriptional and translational downregulation of H-REV107, a class II tumour suppressor gene located on human chromosome 11q11-12. Oncogene 1998, 17:1305-1312.
• [2]Nazarenko I, Schafer R, Sers C: Mechanisms of the HRSL3 tumor suppressor function in ovarian carcinoma cells. J Cell Sci 2007, 120:1393-1404.
• [3]Duncan RE, Sarkadi-Nagy E, Jaworski K, Ahmadian M, Sul HS: Identification and functional characterization of adipose-specific phospholipase A2 (AdPLA). J Biol Chem 2008, 283:25428-25436.
• [4]Hajnal A, Klemenz R, Schafer R: Subtraction cloning of H-rev107, a gene specifically expressed in H-ras resistant fibroblasts. Oncogene 1994, 9:479-490.
• [5]Huang SL, Shyu RY, Yeh MY, Jiang SY: Cloning and characterization of a novel retinoid-inducible gene 1(RIG1) deriving from human gastric cancer cells. Mol Cell Endocrinol 2000, 159:15-24.
• [6]Uyama T, Jin XH, Tsuboi K, Tonai T, Ueda N: Characterization of the human tumor suppressors TIG3 and HRASLS2 as phospholipid-metabolizing enzymes. Biochim Biophys Acta 2009, 1791:1114-1124.
• [7]Shyu RY, Hsieh YC, Tsai FM, Wu CC, Jiang SY: Cloning and functional characterization of the HRASLS2 gene. Amino Acids 2008, 35:129-137.
• [8]Jin XH, Uyama T, Wang J, Okamoto Y, Tonai T, Ueda N: cDNA cloning and characterization of human and mouse Ca(2+)-independent phosphatidylethanolamine N-acyltransferases. Biochim Biophys Acta 2009, 1791:32-38.
• [9]Akiyama H, Hiraki Y, Noda M, Shigeno C, Ito H, Nakamura T: Molecular cloning and biological activity of a novel Ha-Ras suppressor gene predominantly expressed in skeletal muscle, heart, brain, and bone marrow by differential display using clonal mouse EC cells, ATDC5. J Biol Chem 1999, 274:32192-32197.
• [10]Ito H, Akiyama H, Shigeno C, Nakamura T: Isolation, characterization, and chromosome mapping of a human A-C1 Ha-Ras suppressor gene (HRASLS). Cytogenet Cell Genet 2001, 93:36-39.
• [11]Anantharaman V, Aravind L: Evolutionary history, structural features and biochemical diversity of the NlpC/P60 superfamily of enzymes. Genome Biol 2003, 4:R11. BioMed Central Full Text
• [12]Hughes PJ, Stanway G: The 2A proteins of three diverse picornaviruses are related to each other and to the H-rev107 family of proteins involved in the control of cell proliferation. J Gen Virol 2000, 81:201-207.
• [13]Tsai FM, Shyu RY, Jiang SY: RIG1 inhibits the Ras/mitogen-activated protein kinase pathway by suppressing the activation of Ras. Cell Signal 2006, 18:349-358.
• [14]Tsai FM, Shyu RY, Lin SC, Wu CC, Jiang SY: Induction of apoptosis by the retinoid inducible growth regulator RIG1 depends on the NC motif in HtTA cervical cancer cells. BMC Cell Biol 2009, 10:15. BioMed Central Full Text
• [15]Sturniolo MT, Dashti SR, Deucher A, Rorke EA, Broome AM, Chandraratna RA, Keepers T, Eckert RL: A novel tumor suppressor protein promotes keratinocyte terminal differentiation via activation of type I transglutaminase. J Biol Chem 2003, 278:48066-48073.
• [16]Deucher A, Nagpal S, Chandraratna RA, Di Sepio D, Robinson NA, Dashti SR, Eckert RL: The carboxy-terminal hydrophobic domain of TIG3, a class II tumor suppressor protein, is required for appropriate cellular localization and optimal biological activity. Int J Oncol 2000, 17:1195-1203.
• [17]Scharadin TM, Jiang H, Martin S, Eckert RL: TIG3 interaction at the centrosome alters microtubule distribution and centrosome function. J Cell Sci 2012, 125:2604-2614.
• [18]Scharadin TM, Jiang H, Jans R, Rorke EA, Eckert RL: TIG3 tumor suppressor-dependent organelle redistribution and apoptosis in skin cancer cells. PLoS One 2011, 6:e23230.
• [19]Tsai FM, Shyu RY, Jiang SY: RIG1 suppresses Ras activation and induces cellular apoptosis at the Golgi apparatus. Cell Signal 2007, 19:989-999.
• [20]Huang SL, Shyu RY, Yeh MY, Jiang SY: The retinoid-inducible gene I: effect on apoptosis and mitogen-activated kinase signal pathways. Anticancer Res 2002, 22:799-804.
• [21]Sers C, Emmenegger U, Husmann K, Bucher K, Andres AC, Schafer R: Growth-inhibitory activity and downregulation of the class II tumor-suppressor gene H-rev107 in tumor cell lines and experimental tumors. J Cell Biol 1997, 136:935-944.
• [22]Sturniolo MT, Chandraratna RA, Eckert RL: A novel transglutaminase activator forms a complex with type 1 transglutaminase. Oncogene 2005, 24:2963-2972.
• [23]Sers C, Husmann K, Nazarenko I, Reich S, Wiechen K, Zhumabayeva B, Adhikari P, Schroder K, Gontarewicz A, Schafer R: The class II tumour suppressor gene H-REV107-1 is a target of interferon-regulatory factor-1 and is involved in IFNgamma-induced cell death in human ovarian carcinoma cells. Oncogene 2002, 21:2829-2839.
• [24]Uyama T, Morishita J, Jin XH, Okamoto Y, Tsuboi K, Ueda N: The tumor suppressor gene H-Rev107 functions as a novel Ca2 + −independent cytosolic phospholipase A1/2 of the thiol hydrolase type. J Lipid Res 2009, 50:685-693.
• [25]Shinohara N, Uyama T, Jin XH, Tsuboi K, Tonai T, Houchi H, Ueda N: Enzymological analysis of the tumor suppressor A-C1 reveals a novel group of phospholipid-metabolizing enzymes. J Lipid Res 2011, 52:1927-1935.
• [26]Shih TY, Papageorge AG, Stokes PE, Weeks MO, Scolnick EM: Guanine nucleotide-binding and autophosphorylating activities associated with the p21src protein of Harvey murine sarcoma virus. Nature 1980, 287:686-691.
• [27]Bollag G, McCormick F: Regulators and effectors of ras proteins. Annu Rev Cell Biol 1991, 7:601-632.
• [28]Chardin P, Camonis JH, Gale NW, van Aelst L, Schlessinger J, Wigler MH, Bar-Sagi D: Human Sos1: a guanine nucleotide exchange factor for Ras that binds to GRB2. Science 1993, 260:1338-1343.
• [29]Ahearn IM, Haigis K, Bar-Sagi D, Philips MR: Regulating the regulator: post-translational modification of RAS. Nat Rev Mol Cell Biol 2012, 13:39-51.
• [30]Salaun C, Greaves J, Chamberlain LH: The intracellular dynamic of protein palmitoylation. J Biol Chem 2010, 191:1229-1238.
• [31]Choy E, Chiu VK, Silletti J, Feoktistov M, Morimoto T, Michaelson D, Ivanov IE, Philips MR: Endomembrane trafficking of ras: the CAAX motif targets proteins to the ER and Golgi. Cell 1999, 98:69-80.
• [32]Rocks O, Peyker A, Kahms M, Verveer PJ, Koerner C, Lumbierres M, Kuhlmann J, Waldmann H, Wittinghofer A, Bastiaens PI: An acylation cycle regulates localization and activity of palmitoylated Ras isoforms. Science 2005, 307:1746-1752.
• [33]Goodwin JS, Drake KR, Rogers C, Wright L, Lippincott-Schwartz J, Philips MR, Kenworthy AK: Depalmitoylated Ras traffics to and from the Golgi complex via a nonvesicular pathway. J Cell Biol 2005, 170:261-272.
• [34]Misaki R, Morimatsu M, Uemura T, Waguri S, Miyoshi E, Taniguchi N, Matsuda M, Taguchi T: Palmitoylated Ras proteins traffic through recycling endosomes to the plasma membrane during exocytosis. J Cell Biol 2010, 191:23-29.
• [35]Shyu RY, Wu CC, Wang CH, Tsai TC, Wang LK, Chen ML, Jiang SY, Tsai FM: H-rev107 regulates prostaglandin D2 synthase-mediated suppression of cellular invasion in testicular cancer cells. J Biomed Sci 2013, 20:30. BioMed Central Full Text
• [36]Roth AF, Wan J, Bailey AO, Sun B, Kuchar JA, Green WN, Phinney BS, Yates JR 3rd, Davis NG: Global analysis of protein palmitoylation in yeast. Cell 2006, 125:1003-1013.
• [37]Uyama T, Ichi I, Kono N, Inoue A, Tsuboi K, Jin XH, Araki N, Aoki J, Arai H, Ueda N: Regulation of peroxisomal lipid metabolism by catalytic activity of tumor suppressor H-rev107. J Biol Chem 2012, 287:2706-2718.
• [38]Linder ME, Deschenes RJ: Palmitoylation: policing protein stability and traffic. Nat Rev Mol Cell Biol 2007, 8:74-84.
• [39]Dekker FJ, Rocks O, Vartak N, Menninger S, Hedberg C, Balamurugan R, Wetzel S, Renner S, Gerauer M, Scholermann B, Rusch M, Kramer JW, Rauh D, Coates GW, Brunsveld L, Bastiaens PI, Waldmann H: Small-molecule inhibition of APT1 affects Ras localization and signaling. Nat Chem Biol 2010, 6:449-456.
• [40]Dennis EA, Cao J, Hsu YH, Magrioti V, Kokotos G: Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Chem Rev 2011, 111:6130-6185.
• [41]Gelb MH, Jain MK, Berg OG: Inhibition of phospholipase A2. FASEB J 1994, 8:916-924.
• [42]Lucas KK, Dennis EA: The ABC’s of Group IV cytosolic phospholipase A2. Biochim Biophys Acta 2004, 1636:213-218.
• [43]Jenkins CM, Mancuso DJ, Yan W, Sims HF, Gibson B, Gross RW: Identification, cloning, expression, and purification of three novel human calcium-independent phospholipase A2 family members possessing triacylglycerol lipase and acylglycerol transacylase activities. J Biol Chem 2004, 279:48968-48975.
• [44]Eisenberg S, Beckett AJ, Prior IA, Dekker FJ, Hedberg C, Waldmann H, Ehrlich M, Henis YI: Raft protein clustering alters N-Ras membrane interactions and activation pattern. Mol Cell Biol 2011, 31:3938-3952.
• [45]Cox AD: Protein localization: Can too much lipid glue stop Ras? Nat Chem Biol 2010, 6:483-485.
• [46]Wu CC, Shyu RY, Wang CH, Tsai TC, Wang LK, Chen ML, Jiang SY, Tsai FM: Involvement of the prostaglandin D2 signal pathway in retinoid-inducible gene 1 (RIG1)-mediated suppression of cell invasion in testis cancer cells. Biochim Biophys Acta 1823, 2012:2227-2236.
• [47]Reddy ST, Winstead MV, Tischfield JA, Herschman HR: Analysis of the secretory phospholipase A2 that mediates prostaglandin production in mast cells. J Biol Chem 1997, 272:13591-13596.
• [48]Fujishima H, Sanchez Mejia RO, Bingham CO 3rd, Lam BK, Sapirstein A, Bonventre JV, Austen KF, Arm JP: Cytosolic phospholipase A2 is essential for both the immediate and the delayed phases of eicosanoid generation in mouse bone marrow-derived mast cells. Proc Natl Acad Sci U S A 1999, 96:4803-4807.
• [49]Nazarenko I, Kristiansen G, Fonfara S, Guenther R, Gieseler C, Kemmner W, Schafer R, Petersen I, Sers C: H-REV107-1 stimulates growth in non-small cell lung carcinomas via the activation of mitogenic signaling. Am J Pathol 2006, 169:1427-1439.