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BMC Cancer
Published in: BMC Cancer 1/2014

Open Access 01-12-2014 | Research article

Conformational changes and translocation of tissue-transglutaminase to the plasma membranes: role in cancer cell migration

Authors: Ambrish Kumar, Jianjun Hu, Holly A LaVoie, Kenneth B Walsh, Donald J DiPette, Ugra S Singh

Published in: BMC Cancer | Issue 1/2014

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Abstract

Background

Tissue-transglutaminase (TG2), a dual function G-protein, plays key roles in cell differentiation and migration. In our previous studies we reported the mechanism of TG2-induced cell differentiation. In present study, we explored the mechanism of how TG2 may be involved in cell migration.

Methods

To study the mechanism of TG2-mediated cell migration, we used neuroblastoma cells (SH-SY5Y) which do not express TG2, neuroblastoma cells expressing exogenous TG2 (SHYTG2), and pancreatic cancer cells which express high levels of endogenous TG2. Resveratrol, a natural compound previously shown to inhibit neuroblastoma and pancreatic cancer in the animal models, was utilized to investigate the role of TG2 in cancer cell migration. Immunofluorescence assays were employed to detect expression and intracellular localization of TG2, and calcium levels in the migrating cells. Native gel electrophoresis was performed to analyze resveratrol-induced cellular distribution and conformational states of TG2 in migrating cells. Data are presented as the mean and standard deviation of at least 3 independent experiments. Comparisons were made among groups using one-way ANOVA followed by Tukey-Kramer ad hoc test.

Results

TG2 containing cells (SHYTG2 and pancreatic cancer cells) exhibit increased cell migration and invasion in collagen-coated and matrigel-coated transwell plate assays, respectively. Resveratrol (1 μM-10 μM) prevented migration of TG2-expressing cells. During the course of migration, resveratrol increased the immunoreactivity of TG2 without affecting the total TG2 protein level in migrating cells. In these cells, resveratrol increased calcium levels, and depletion of intracellular calcium by a calcium chelator, BAPTA, attenuated resveratrol-enhanced TG2 immunoreactivity. In native-polyacrylamide gels, we detected an additional TG2 protein band with slower migration in total cell lysates of resveratrol treated cells. This TG2 form is non-phosphorylated, exclusively present in plasma membrane fractions and sensitive to intracellular Ca2+ concentration suggesting a calcium requirement in TG2-regulated cell migration.

Conclusions

Taken together, we conclude that resveratrol induces conformational changes in TG2, and that Ca2+-mediated TG2 association with the plasma membrane is responsible for the inhibitory effects of resveratrol on cell migration.
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Literature
1.
Folk JE, Finlayson JS: The epsilon-(gamma-glutamyl) lysine crosslink and the catalytic role of transglutaminases. Adv Protein Chem. 1977, 31: 1-133.CrossRefPubMed
2.
Lorand L, Graham RM: Transglutaminases: crosslinking enzymes with pleiotropic functions. Nat Rev Mol Cell Biol. 2003, 4 (2): 140-156. 10.1038/nrm1014.CrossRefPubMed
3.
Singh US, Erickson JW, Cerione RA: Identification and biochemical characterization of an 80 kilodalton GTP-binding/transglutaminase from rabbit liver nuclei. Biochemistry. 1995, 34 (48): 15863-15871. 10.1021/bi00048a032.CrossRefPubMed
4.
Nakaoka H, Perez DM, Baek KJ, Das T, Husain A, Misono K, Im MJ, Graham RM: Gh: a GTP-binding protein with transglutaminase activity and receptor signaling function. Science. 1994, 264 (5165): 1593-1596. 10.1126/science.7911253.CrossRefPubMed
5.
Begg GE, Carrington L, Stokes PH, Matthews JM, Wouters MA, Husain A, Lorand L, Iismaa SE, Graham RM: Mechanism of allosteric regulation of transglutaminase 2 by GTP. Proc Natl Acad Sci U S A. 2006, 103 (52): 19683-19688. 10.1073/pnas.0609283103.CrossRefPubMedPubMedCentral
6.
Telci D, Wang Z, Li X, Verderio EA, Humphries MJ, Baccarini M, Basaga H, Griffin M: Fibronectin-tissue transglutaminase matrix rescues RGD-impaired cell adhesion through syndecan-4 and beta1 integrin co-signaling. J Biol Chem. 2008, 283 (30): 20937-20947. 10.1074/jbc.M801763200.CrossRefPubMedPubMedCentral
7.
Collighan RJ, Griffin M: Transglutaminase 2 cross-linking of matrix proteins: biological significance and medical applications. Amino Acids. 2009, 36 (4): 659-670. 10.1007/s00726-008-0190-y.CrossRefPubMed
8.
Akimov SS, Belkin AM: Cell-surface transglutaminase promotes fibronectin assembly via interaction with the gelatin-binding domain of fibronectin: a role in TGFbeta-dependent matrix deposition. J Cell Sci. 2001, 114 (Pt 16): 2989-3000.PubMed
9.
Balklava Z, Verderio E, Collighan R, Gross S, Adams J, Griffin M: Analysis of tissue transglutaminase function in the migration of Swiss 3 T3 fibroblasts: the active-state conformation of the enzyme does not affect cell motility but is important for its secretion. J Biol Chem. 2002, 277 (19): 16567-16575. 10.1074/jbc.M109836200.CrossRefPubMed
10.
Lin CY, Tsai PH, Kandaswami CC, Chang GD, Cheng CH, Huang CJ, Lee PP, Hwang JJ, Lee MT: Role of tissue transglutaminase 2 in the acquisition of a mesenchymal-like phenotype in highly invasive A431 tumor cells. Mol Cancer. 2011, 10: 87-10.1186/1476-4598-10-87.CrossRefPubMedPubMedCentral
11.
Mehta K, Kumar A, Kim HI: Transglutaminase 2: a multi-tasking protein in the complex circuitry of inflammation and cancer. Biochem Pharmacol. 2010, 80 (12): 1921-1929. 10.1016/j.bcp.2010.06.029.CrossRefPubMed
12.
Verma A, Wang H, Manavathi B, Fok JY, Mann AP, Kumar R, Mehta K: Increased expression of tissue transglutaminase in pancreatic ductal adenocarcinoma and its implications in drug resistance and metastasis. Cancer Res. 2006, 66 (21): 10525-10533. 10.1158/0008-5472.CAN-06-2387.CrossRefPubMed
13.
Hwang JY, Mangala LS, Fok JY, Lin YG, Merritt WM, Spannuth WA, Nick AM, Fiterman DJ, Vivas-Mejia PE, Deavers MT, Coleman RL, Lopez-Berestein G, Mehta K, Sood AK: Clinical and biological significance of tissue transglutaminase in ovarian carcinoma. Cancer Res. 2008, 68 (14): 5849-5858. 10.1158/0008-5472.CAN-07-6130.CrossRefPubMedPubMedCentral
14.
Satpathy M, Cao L, Pincheira R, Emerson R, Bigsby R, Nakshatri H, Matei D: Enhanced peritoneal ovarian tumor dissemination by tissue transglutaminase. Cancer Res. 2007, 67 (15): 7194-7202. 10.1158/0008-5472.CAN-07-0307.CrossRefPubMed
15.
Mangala LS, Fok JY, Zorrilla-Calancha IR, Verma A, Mehta K: Tissue transglutaminase expression promotes cell attachment, invasion and survival in breast cancer cells. Oncogene. 2007, 26 (17): 2459-2470. 10.1038/sj.onc.1210035.CrossRefPubMed
16.
Fok JY, Ekmekcioglu S, Mehta K: Implications of tissue transglutaminase expression in malignant melanoma. Mol Cancer Ther. 2006, 5 (6): 1493-1503. 10.1158/1535-7163.MCT-06-0083.CrossRefPubMed
17.
Shao M, Cao L, Shen C, Satpathy M, Chelladurai B, Bigsby RM, Nakshatri H, Matei D: Epithelial-to-mesenchymal transition and ovarian tumor progression induced by tissue transglutaminase. Cancer Res. 2009, 69 (24): 9192-9201. 10.1158/0008-5472.CAN-09-1257.CrossRefPubMed
18.
Kumar A, Xu J, Brady S, Gao H, Yu D, Reuben J, Mehta K: Tissue transglutaminase promotes drug resistance and invasion by inducing mesenchymal transition in mammary epithelial cells. PLoS One. 2010, 5 (10): e13390-10.1371/journal.pone.0013390.CrossRefPubMedPubMedCentral
19.
Verma A, Guha S, Diagaradjane P, Kunnumakkara AB, Sanguino AM, Lopez-Berestein G, Sood AK, Aggarwal BB, Krishnan S, Gelovani JG, Mehta K: Therapeutic significance of elevated tissue transglutaminase expression in pancreatic cancer. Clin Cancer Res. 2008, 14 (8): 2476-2483. 10.1158/1078-0432.CCR-07-4529.CrossRefPubMed
20.
Antonyak MA, Li B, Regan AD, Feng Q, Dusaban SS, Cerione RA: Tissue transglutaminase is an essential participant in the epidermal growth factor-stimulated signaling pathway leading to cancer cell migration and invasion. J Biol Chem. 2009, 284 (27): 17914-17925. 10.1074/jbc.M109.013037.CrossRefPubMedPubMedCentral
21.
Singh US, Pan J, Kao YL, Joshi S, Young KL, Baker KM: Tissue Transglutaminase Mediates Activation of RhoA and MAP Kinase Pathways during Retinoic Acid-induced Neuronal Differentiation of SH-SY5Y Cells. J Biol Chem. 2003, 278 (1): 391-399.CrossRefPubMed
22.
Singh US, Kunar MT, Kao YL, Baker KM: Role of transglutaminase II in retinoic acid-induced activation of RhoA- associated kinase-2. EMBO J. 2001, 20 (10): 2413-2423. 10.1093/emboj/20.10.2413.CrossRefPubMedPubMedCentral
23.
Roy SK, Chen Q, Fu J, Shankar S, Srivastava RK: Resveratrol inhibits growth of orthotopic pancreatic tumors through activation of FOXO transcription factors. PLoS One. 2011, 6 (9): e25166-10.1371/journal.pone.0025166.CrossRefPubMedPubMedCentral
24.
van Ginkel PR, Sareen D, Subramanian L, Walker Q, Darjatmoko SR, Lindstrom MJ, Kulkarni A, Albert DM, Polans AS: Resveratrol inhibits tumor growth of human neuroblastoma and mediates apoptosis by directly targeting mitochondria. Clin Cancer Res. 2007, 13 (17): 5162-5169. 10.1158/1078-0432.CCR-07-0347.CrossRefPubMed
25.
Bishayee A: Cancer prevention and treatment with resveratrol: from rodent studies to clinical trials. Cancer Prev Res (Phila). 2009, 2 (5): 409-418. 10.1158/1940-6207.CAPR-08-0160.CrossRef
26.
Chen Y, Tseng SH, Lai HS, Chen WJ: Resveratrol-induced cellular apoptosis and cell cycle arrest in neuroblastoma cells and antitumor effects on neuroblastoma in mice. Surgery. 2004, 136 (1): 57-66. 10.1016/j.surg.2004.01.017.CrossRefPubMed
27.
Pervaiz S, Holme AL: Resveratrol: its biologic targets and functional activity. Antioxid Redox Signal. 2009, 11 (11): 2851-2897. 10.1089/ars.2008.2412.CrossRefPubMed
28.
Joshi S, Guleria R, Pan J, DiPette D, Singh US: Retinoic acid receptors and tissue-transglutaminase mediate short-term effect of retinoic acid on migration and invasion of neuroblastoma SH-SY5Y cells. Oncogene. 2006, 25 (2): 240-247.PubMed
29.
Kumar A, Singh CK, DiPette DD, Singh US: Ethanol impairs activation of retinoic acid receptors in cerebellar granule cells in a rodent model of fetal alcohol spectrum disorders. Alcohol Clin Exp Res. 2010, 34 (5): 928-937. 10.1111/j.1530-0277.2010.01166.x.CrossRefPubMedPubMedCentral
30.
Kumar A, Singh CK, Lavoie HA, Dipette DJ, Singh US: Resveratrol restores Nrf2 level and prevents ethanol-induced toxic effects in the cerebellum of a rodent model of fetal alcohol spectrum disorders. Mol Pharmacol. 2011, 80 (3): 446-457. 10.1124/mol.111.071126.CrossRefPubMedPubMedCentral
31.
Singh US, Cerione RA: Biochemical effects of retinoic acid on GTP-binding Protein/Transglutaminases in HeLa cells. Stimulation of GTP-binding and transglutaminase activity, membrane association, and phosphatidylinositol lipid turnover. J Biol Chem. 1996, 271 (44): 27292-27298. 10.1074/jbc.271.44.27292.CrossRefPubMed
32.
Tucholski J, Lesort M, Johnson GV: Tissue transglutaminase is essential for neurite outgrowth in human neuroblastoma SH-SY5Y cells. Neuroscience. 2001, 102 (2): 481-491. 10.1016/S0306-4522(00)00482-6.CrossRefPubMed
33.
Mishra S, Murphy LJ: Tissue transglutaminase has intrinsic kinase activity: identification of transglutaminase 2 as an insulin-like growth factor-binding protein-3 kinase. J Biol Chem. 2004, 279 (23): 23863-23868. 10.1074/jbc.M311919200.CrossRefPubMed
34.
Pinkas DM, Strop P, Brunger AT, Khosla C: Transglutaminase 2 undergoes a large conformational change upon activation. PLoS Biol. 2007, 5 (12): e327-10.1371/journal.pbio.0050327.CrossRefPubMedPubMedCentral
35.
Park SJ, Ahmad F, Philp A, Baar K, Williams T, Luo H, Ke H, Rehmann H, Taussig R, Brown AL, Kim MK, Beaven MA, Burgin AB, Manganiello V, Chung JH: Resveratrol ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases. Cell. 2012, 148 (3): 421-433. 10.1016/j.cell.2012.01.017.CrossRefPubMedPubMedCentral
36.
Mi LY, Ettenson DS, Edelman ER: Phospholipase C-delta extends intercellular signalling range and responses to injury-released growth factors in non-excitable cells. Cell Prolif. 2008, 41 (4): 671-690. 10.1111/j.1365-2184.2008.00544.x.CrossRefPubMedPubMedCentral
37.
Padin JF, de Diego AM, Fernandez-Morales JC, Merino C, Maroto M, Calvo-Gallardo E, Arranz JA, Yanez M, Garcia AG: Resveratrol augments nitric oxide generation and causes store calcium release in chromaffin cells. Eur J Pharmacol. 2012, 685 (1–3): 99-107.CrossRefPubMed
38.
Stamnaes J, Pinkas DM, Fleckenstein B, Khosla C, Sollid LM: Redox regulation of transglutaminase 2 activity. J Biol Chem. 2010, 285 (33): 25402-25409. 10.1074/jbc.M109.097162.CrossRefPubMedPubMedCentral
39.
Kiraly R, Csosz E, Kurtan T, Antus S, Szigeti K, Simon-Vecsei Z, Korponay-Szabo IR, Keresztessy Z, Fesus L: Functional significance of five noncanonical Ca2 + −binding sites of human transglutaminase 2 characterized by site-directed mutagenesis. FEBS J. 2009, 276 (23): 7083-7096. 10.1111/j.1742-4658.2009.07420.x.CrossRefPubMed
40.
Bergamini CM: GTP modulates calcium binding and cation-induced conformational changes in erythrocyte transglutaminase. FEBS Lett. 1988, 239 (2): 255-258. 10.1016/0014-5793(88)80928-1.CrossRefPubMed
41.
Kiraly R, Demeny M, Fesus L: Protein transamidation by transglutaminase 2 in cells: a disputed Ca2 + −dependent action of a multifunctional protein. FEBS J. 2011, 278 (24): 4717-4739. 10.1111/j.1742-4658.2011.08345.x.CrossRefPubMed
42.
Liu S, Cerione RA, Clardy J: Structural basis for the guanine nucleotide-binding activity of tissue transglutaminase and its regulation of transamidation activity. Proc Natl Acad Sci U S A. 2002, 99 (5): 2743-2747. 10.1073/pnas.042454899.CrossRefPubMedPubMedCentral
43.
Di Venere A, Rossi A, De Matteis F, Rosato N, Agro AF, Mei G: Opposite effects of Ca (2+) and GTP binding on tissue transglutaminase tertiary structure. J Biol Chem. 2000, 275 (6): 3915-3921. 10.1074/jbc.275.6.3915.CrossRefPubMed
44.
Casadio R, Polverini E, Mariani P, Spinozzi F, Carsughi F, Fontana A, Polverino de Laureto P, Matteucci G, Bergamini CM: The structural basis for the regulation of tissue transglutaminase by calcium ions. Eur J Biochem. 1999, 262 (3): 672-679. 10.1046/j.1432-1327.1999.00437.x.CrossRefPubMed
45.
Boroughs LK, Antonyak MA, Johnson JL, Cerione RA: A unique role for heat shock protein 70 and its binding partner tissue transglutaminase in cancer cell migration. J Biol Chem. 2011, 286 (43): 37094-37107. 10.1074/jbc.M111.242438.CrossRefPubMedPubMedCentral
46.
Lai TS, Davies C, Greenberg CS: Human tissue transglutaminase is inhibited by pharmacologic and chemical acetylation. Protein Sci. 2010, 19 (2): 229-235. 10.1002/pro.301.CrossRefPubMed
47.
Jeong EM, Kim CW, Cho SY, Jang GY, Shin DM, Jeon JH, Kim IG: Degradation of transglutaminase 2 by calcium-mediated ubiquitination responding to high oxidative stress. FEBS Lett. 2009, 583 (4): 648-654. 10.1016/j.febslet.2009.01.032.CrossRefPubMed
48.
Park D, Choi SS, Ha KS: Transglutaminase 2: a multi-functional protein in multiple subcellular compartments. Amino Acids. 2010, 39 (3): 619-631. 10.1007/s00726-010-0500-z.CrossRefPubMed
49.
Belkin AM, Tsurupa G, Zemskov E, Veklich Y, Weisel JW, Medved L: Transglutaminase-mediated oligomerization of the fibrin (ogen) alphaC domains promotes integrin-dependent cell adhesion and signaling. Blood. 2005, 105 (9): 3561-3568. 10.1182/blood-2004-10-4089.CrossRefPubMedPubMedCentral
50.
Satpathy M, Shao M, Emerson R, Donner DB, Matei D: Tissue transglutaminase regulates matrix metalloproteinase-2 in ovarian cancer by modulating cAMP-response element-binding protein activity. J Biol Chem. 2009, 284 (23): 15390-15399. 10.1074/jbc.M808331200.CrossRefPubMedPubMedCentral
51.
Deryugina EI, Quigley JP: Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev. 2006, 25 (1): 9-34. 10.1007/s10555-006-7886-9.CrossRefPubMed
52.
Borra MT, Smith BC, Denu JM: Mechanism of human SIRT1 activation by resveratrol. J Biol Chem. 2005, 280 (17): 17187-17195. 10.1074/jbc.M501250200.CrossRefPubMed
53.
Wang Z, Griffin M: The role of TG2 in regulating S100A4-mediated mammary tumour cell migration. PLoS One. 2013, 8 (3): e57017-10.1371/journal.pone.0057017.CrossRefPubMedPubMedCentral
54.
Timmers S, Konings E, Bilet L, Houtkooper RH, van de Weijer T, Goossens GH, Hoeks J, van der Krieken S, Ryu D, Kersten S, Moonen-Kornips E, Hesselink MK, Kunz I, Schrauwen-Hinderling VB, Blaak EE, Auwerx J, Schrauwen P: Calorie restriction-like effects of 30 days of resveratrol supplementation on energy metabolism and metabolic profile in obese humans. Cell Metab. 2011, 14 (5): 612-622. 10.1016/j.cmet.2011.10.002.CrossRefPubMed
55.
Patel KR, Andreadi C, Britton RG, Horner-Glister E, Karmokar A, Sale S, Brown VA, Brenner DE, Singh R, Steward WP, Gescher AJ, Brown K: Sulfate metabolites provide an intracellular pool for resveratrol generation and induce autophagy with senescence. Sci Transl Med. 2013, 5 (205): 205ra133-CrossRefPubMed
Metadata
Title
Conformational changes and translocation of tissue-transglutaminase to the plasma membranes: role in cancer cell migration
Authors
Ambrish Kumar
Jianjun Hu
Holly A LaVoie
Kenneth B Walsh
Donald J DiPette
Ugra S Singh
Publication date
01-12-2014
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2014
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-14-256

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