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. 1992 Oct 2;119(2):463–474. doi: 10.1083/jcb.119.2.463

Expression of tissue transglutaminase in Balb-C 3T3 fibroblasts: effects on cellular morphology and adhesion

PMCID: PMC2289646  PMID: 1356992

Abstract

Tissue transglutaminase is a cytosolic enzyme whose primary function is to catalyze the covalent cross-linking of proteins. To investigate the functions of this enzyme in physiological systems, we have established lines of Balb-C 3T3 fibroblasts stably transfected with a constitutive tissue transglutaminase expression plasmid. Several cell lines expressing high levels of catalytically active tissue transglutaminase have been isolated and characterized. Transglutaminase-transfected cells showed morphologic features quite distinct from their nontransfected counterparts. Many of the cells showed an extended and very flattened morphology that reflected increased adhesion of the cells to the substratum. Other cells, particularly those showing the highest levels of intracellular transglutaminase expression, showed extensive membrane blebbing and cellular fragmentation. The results of these experiments suggest that the induction and activation of tissue transglutaminase may contribute both to changes in cellular morphology and adhesiveness.

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Selected References

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  1. Achyuthan K. E., Greenberg C. S. Identification of a guanosine triphosphate-binding site on guinea pig liver transglutaminase. Role of GTP and calcium ions in modulating activity. J Biol Chem. 1987 Feb 5;262(4):1901–1906. [PubMed] [Google Scholar]
  2. Aeschlimann D., Paulsson M. Cross-linking of laminin-nidogen complexes by tissue transglutaminase. A novel mechanism for basement membrane stabilization. J Biol Chem. 1991 Aug 15;266(23):15308–15317. [PubMed] [Google Scholar]
  3. Arends M. J., Wyllie A. H. Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol. 1991;32:223–254. doi: 10.1016/b978-0-12-364932-4.50010-1. [DOI] [PubMed] [Google Scholar]
  4. Bowness J. M., Folk J. E., Timpl R. Identification of a substrate site for liver transglutaminase on the aminopropeptide of type III collagen. J Biol Chem. 1987 Jan 25;262(3):1022–1024. [PubMed] [Google Scholar]
  5. Byrd J. C., Lichti U. Two types of transglutaminase in the PC12 pheochromocytoma cell line. Stimulation by sodium butyrate. J Biol Chem. 1987 Aug 25;262(24):11699–11705. [PubMed] [Google Scholar]
  6. Cai D., Ben T., De Luca L. M. Retinoids induce tissue transglutaminase in NIH-3T3 cells. Biochem Biophys Res Commun. 1991 Mar 29;175(3):1119–1124. doi: 10.1016/0006-291x(91)91681-2. [DOI] [PubMed] [Google Scholar]
  7. Chiocca E. A., Davies P. J., Stein J. P. The molecular basis of retinoic acid action. Transcriptional regulation of tissue transglutaminase gene expression in macrophages. J Biol Chem. 1988 Aug 15;263(23):11584–11589. [PubMed] [Google Scholar]
  8. Cornwell M. M., Juliano R. L., Davies P. J. Inhibition of the adhesion of Chinese hamster ovary cells by the naphthylsulfonamides dansylcadaverine and N-(6-aminohexyl)-5-chloro-1-naphthylenesulfonamide (W7). Biochim Biophys Acta. 1983 Jun 2;762(3):414–419. doi: 10.1016/0167-4889(83)90006-x. [DOI] [PubMed] [Google Scholar]
  9. Cullen B. R. Use of eukaryotic expression technology in the functional analysis of cloned genes. Methods Enzymol. 1987;152:684–704. doi: 10.1016/0076-6879(87)52074-2. [DOI] [PubMed] [Google Scholar]
  10. Davies P. J., Chiocca E. A., Basilion J. P., Poddar S., Stein J. P. Transglutaminases and their regulation: implications for polyamine metabolism. Adv Exp Med Biol. 1988;250:391–401. doi: 10.1007/978-1-4684-5637-0_34. [DOI] [PubMed] [Google Scholar]
  11. Davies P. J., Cornwell M. M., Johnson J. D., Reggianni A., Myers M., Murtaugh M. P. Studies on the effects of dansylcadaverine and related compounds on receptor-mediated endocytosis in cultured cells. Diabetes Care. 1984 May-Jun;7 (Suppl 1):35–41. [PubMed] [Google Scholar]
  12. Davies P. J., Murtaugh M. P., Moore W. T., Jr, Johnson G. S., Lucas D. Retinoic acid-induced expression of tissue transglutaminase in human promyelocytic leukemia (HL-60) cells. J Biol Chem. 1985 Apr 25;260(8):5166–5174. [PubMed] [Google Scholar]
  13. De Luca L. M., Adamo S., Kato S. Retinoids and cell adhesion. Methods Enzymol. 1990;190:81–91. doi: 10.1016/0076-6879(90)90012-p. [DOI] [PubMed] [Google Scholar]
  14. Eghbali B., Kessler J. A., Spray D. C. Expression of gap junction channels in communication-incompetent cells after stable transfection with cDNA encoding connexin 32. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1328–1331. doi: 10.1073/pnas.87.4.1328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fesus L., Davies P. J., Piacentini M. Apoptosis: molecular mechanisms in programmed cell death. Eur J Cell Biol. 1991 Dec;56(2):170–177. [PubMed] [Google Scholar]
  16. Fesus L., Metsis M. L., Muszbek L., Koteliansky V. E. Transglutaminase-sensitive glutamine residues of human plasma fibronectin revealed by studying its proteolytic fragments. Eur J Biochem. 1986 Jan 15;154(2):371–374. doi: 10.1111/j.1432-1033.1986.tb09407.x. [DOI] [PubMed] [Google Scholar]
  17. Fesus L., Szucs E. F., Barrett K. E., Metcalfe D. D., Folk J. E. Activation of transglutaminase and production of protein-bound gamma-glutamylhistamine in stimulated mouse mast cells. J Biol Chem. 1985 Nov 5;260(25):13771–13778. [PubMed] [Google Scholar]
  18. Fesus L., Tarcsa E. Formation of N epsilon-(gamma-glutamyl)-lysine isodipeptide in Chinese-hamster ovary cells. Biochem J. 1989 Nov 1;263(3):843–848. doi: 10.1042/bj2630843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fesus L., Thomazy V., Autuori F., Ceru M. P., Tarcsa E., Piacentini M. Apoptotic hepatocytes become insoluble in detergents and chaotropic agents as a result of transglutaminase action. FEBS Lett. 1989 Mar 13;245(1-2):150–154. doi: 10.1016/0014-5793(89)80210-8. [DOI] [PubMed] [Google Scholar]
  20. Fesus L., Thomazy V., Falus A. Induction and activation of tissue transglutaminase during programmed cell death. FEBS Lett. 1987 Nov 16;224(1):104–108. doi: 10.1016/0014-5793(87)80430-1. [DOI] [PubMed] [Google Scholar]
  21. Fesus L., Thomazy V. Searching for the function of tissue transglutaminase: its possible involvement in the biochemical pathway of programmed cell death. Adv Exp Med Biol. 1988;231:119–134. doi: 10.1007/978-1-4684-9042-8_10. [DOI] [PubMed] [Google Scholar]
  22. Folk J. E. Mechanism and basis for specificity of transglutaminase-catalyzed epsilon-(gamma-glutamyl) lysine bond formation. Adv Enzymol Relat Areas Mol Biol. 1983;54:1–56. doi: 10.1002/9780470122990.ch1. [DOI] [PubMed] [Google Scholar]
  23. Folk J. E., Park M. H., Chung S. I., Schrode J., Lester E. P., Cooper H. L. Polyamines as physiological substrates for transglutaminases. J Biol Chem. 1980 Apr 25;255(8):3695–3700. [PubMed] [Google Scholar]
  24. Folk J. E. Transglutaminases. Annu Rev Biochem. 1980;49:517–531. doi: 10.1146/annurev.bi.49.070180.002505. [DOI] [PubMed] [Google Scholar]
  25. Gentile V., Saydak M., Chiocca E. A., Akande O., Birckbichler P. J., Lee K. N., Stein J. P., Davies P. J. Isolation and characterization of cDNA clones to mouse macrophage and human endothelial cell tissue transglutaminases. J Biol Chem. 1991 Jan 5;266(1):478–483. [PubMed] [Google Scholar]
  26. Greenberg C. S., Achyuthan K. E., Borowitz M. J., Shuman M. A. The transglutaminase in vascular cells and tissues could provide an alternate pathway for fibrin stabilization. Blood. 1987 Sep;70(3):702–709. [PubMed] [Google Scholar]
  27. Greenberg C. S., Birckbichler P. J., Rice R. H. Transglutaminases: multifunctional cross-linking enzymes that stabilize tissues. FASEB J. 1991 Dec;5(15):3071–3077. doi: 10.1096/fasebj.5.15.1683845. [DOI] [PubMed] [Google Scholar]
  28. Grundmann U., Amann E., Zettlmeissl G., Küpper H. A. Characterization of cDNA coding for human factor XIIIa. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8024–8028. doi: 10.1073/pnas.83.21.8024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hand D., Bungay P. J., Elliott B. M., Griffin M. Activation of transglutaminase at calcium levels consistent with a role for this enzyme as a calcium receptor protein. Biosci Rep. 1985 Dec;5(12):1079–1086. doi: 10.1007/BF01119629. [DOI] [PubMed] [Google Scholar]
  30. Ichinose A., Bottenus R. E., Davie E. W. Structure of transglutaminases. J Biol Chem. 1990 Aug 15;265(23):13411–13414. [PubMed] [Google Scholar]
  31. Ichinose A., Hendrickson L. E., Fujikawa K., Davie E. W. Amino acid sequence of the a subunit of human factor XIII. Biochemistry. 1986 Nov 4;25(22):6900–6906. doi: 10.1021/bi00370a025. [DOI] [PubMed] [Google Scholar]
  32. Ikura K., Nasu T., Yokota H., Tsuchiya Y., Sasaki R., Chiba H. Amino acid sequence of guinea pig liver transglutaminase from its cDNA sequence. Biochemistry. 1988 Apr 19;27(8):2898–2905. doi: 10.1021/bi00408a035. [DOI] [PubMed] [Google Scholar]
  33. Jiang S. Y., Jordan V. C. Growth regulation of estrogen receptor-negative breast cancer cells transfected with complementary DNAs for estrogen receptor. J Natl Cancer Inst. 1992 Apr 15;84(8):580–591. doi: 10.1093/jnci/84.8.580. [DOI] [PubMed] [Google Scholar]
  34. Juprelle-Soret M., Wattiaux-De Coninck S., Wattiaux R. Subcellular localization of transglutaminase. Effect of collagen. Biochem J. 1988 Mar 1;250(2):421–427. doi: 10.1042/bj2500421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Keown W. A., Campbell C. R., Kucherlapati R. S. Methods for introducing DNA into mammalian cells. Methods Enzymol. 1990;185:527–537. doi: 10.1016/0076-6879(90)85043-n. [DOI] [PubMed] [Google Scholar]
  36. Kim H. C., Idler W. W., Kim I. G., Han J. H., Chung S. I., Steinert P. M. The complete amino acid sequence of the human transglutaminase K enzyme deduced from the nucleic acid sequences of cDNA clones. J Biol Chem. 1991 Jan 5;266(1):536–539. [PubMed] [Google Scholar]
  37. Kim H. C., Lewis M. S., Gorman J. J., Park S. C., Girard J. E., Folk J. E., Chung S. I. Protransglutaminase E from guinea pig skin. Isolation and partial characterization. J Biol Chem. 1990 Dec 15;265(35):21971–21978. [PubMed] [Google Scholar]
  38. Kinsella M. G., Wight T. N. Formation of high molecular weight dermatan sulfate proteoglycan in bovine aortic endothelial cell cultures. Evidence for transglutaminase-catalyzed cross-linking to fibronectin. J Biol Chem. 1990 Oct 15;265(29):17891–17898. [PubMed] [Google Scholar]
  39. Kinsella M. G., Wight T. N. Modulation of sulfated proteoglycan synthesis by bovine aortic endothelial cells during migration. J Cell Biol. 1986 Mar;102(3):679–687. doi: 10.1083/jcb.102.3.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Korner G., Schneider D. E., Purdon M. A., Bjornsson T. D. Bovine aortic endothelial cell transglutaminase. Enzyme characterization and regulation of activity. Biochem J. 1989 Sep 1;262(2):633–641. doi: 10.1042/bj2620633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Lee K. N., Birckbichler P. J., Patterson M. K., Jr, Conway E., Maxwell M. Induction of cellular transglutaminase biosynthesis by sodium butyrate. Biochim Biophys Acta. 1987 Apr 2;928(1):22–28. doi: 10.1016/0167-4889(87)90081-4. [DOI] [PubMed] [Google Scholar]
  42. Lee K. N., Fesus L., Yancey S. T., Girard J. E., Chung S. I. Development of selective inhibitors of transglutaminase. Phenylthiourea derivatives. J Biol Chem. 1985 Nov 25;260(27):14689–14694. [PubMed] [Google Scholar]
  43. Lorand L., Conrad S. M. Transglutaminases. Mol Cell Biochem. 1984;58(1-2):9–35. doi: 10.1007/BF00240602. [DOI] [PubMed] [Google Scholar]
  44. Lorand L. Fibrinoligase: the fibrin-stabilizing factor system of blood plasma. Ann N Y Acad Sci. 1972 Dec 8;202:6–30. doi: 10.1111/j.1749-6632.1972.tb16319.x. [DOI] [PubMed] [Google Scholar]
  45. Martinez J., Rich E., Barsigian C. Transglutaminase-mediated cross-linking of fibrinogen by human umbilical vein endothelial cells. J Biol Chem. 1989 Dec 5;264(34):20502–20508. [PubMed] [Google Scholar]
  46. Moore W. T., Jr, Murtaugh M. P., Davies P. J. Retinoic acid-induced expression of tissue transglutaminase in mouse peritoneal macrophages. J Biol Chem. 1984 Oct 25;259(20):12794–12802. [PubMed] [Google Scholar]
  47. Murtaugh M. P., Mehta K., Johnson J., Myers M., Juliano R. L., Davies P. J. Induction of tissue transglutaminase in mouse peritoneal macrophages. J Biol Chem. 1983 Sep 25;258(18):11074–11081. [PubMed] [Google Scholar]
  48. Nara K., Nakanishi K., Hagiwara H., Wakita K., Kojima S., Hirose S. Retinol-induced morphological changes of cultured bovine endothelial cells are accompanied by a marked increase in transglutaminase. J Biol Chem. 1989 Nov 15;264(32):19308–19312. [PubMed] [Google Scholar]
  49. Orrenius S., McConkey D. J., Bellomo G., Nicotera P. Role of Ca2+ in toxic cell killing. Trends Pharmacol Sci. 1989 Jul;10(7):281–285. doi: 10.1016/0165-6147(89)90029-1. [DOI] [PubMed] [Google Scholar]
  50. Paye M., Lapière C. M. The lack of attachment of transformed embryonic lung epithelial cells to collagen I is corrected by fibronectin and FXIII. J Cell Sci. 1986 Dec;86:95–107. doi: 10.1242/jcs.86.1.95. [DOI] [PubMed] [Google Scholar]
  51. Phillips M. A., Stewart B. E., Qin Q., Chakravarty R., Floyd E. E., Jetten A. M., Rice R. H. Primary structure of keratinocyte transglutaminase. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9333–9337. doi: 10.1073/pnas.87.23.9333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Piacentini M., Autuori F., Dini L., Farrace M. G., Ghibelli L., Piredda L., Fesus L. "Tissue" transglutaminase is specifically expressed in neonatal rat liver cells undergoing apoptosis upon epidermal growth factor-stimulation. Cell Tissue Res. 1991 Feb;263(2):227–235. doi: 10.1007/BF00318764. [DOI] [PubMed] [Google Scholar]
  53. Piacentini M., Martinet N., Beninati S., Folk J. E. Free and protein-conjugated polyamines in mouse epidermal cells. Effect of high calcium and retinoic acid. J Biol Chem. 1988 Mar 15;263(8):3790–3794. [PubMed] [Google Scholar]
  54. Piazza G. A., Callanan H. M., Mowery J., Hixson D. C. Evidence for a role of dipeptidyl peptidase IV in fibronectin-mediated interactions of hepatocytes with extracellular matrix. Biochem J. 1989 Aug 15;262(1):327–334. doi: 10.1042/bj2620327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Rice R. H., Green H. Relation of protein synthesis and transglutaminase activity to formation of the cross-linked envelope during terminal differentiation of the cultured human epidermal keratinocyte. J Cell Biol. 1978 Mar;76(3):705–711. doi: 10.1083/jcb.76.3.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Rouault T. A., Hentze M. W., Dancis A., Caughman W., Harford J. B., Klausner R. D. Influence of altered transcription on the translational control of human ferritin expression. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6335–6339. doi: 10.1073/pnas.84.18.6335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Sane D. C., Moser T. L., Greenberg C. S. Vitronectin in the substratum of endothelial cells is cross-linked and phosphorylated. Biochem Biophys Res Commun. 1991 Jan 31;174(2):465–469. doi: 10.1016/0006-291x(91)91439-j. [DOI] [PubMed] [Google Scholar]
  58. Sane D. C., Moser T. L., Pippen A. M., Parker C. J., Achyuthan K. E., Greenberg C. S. Vitronectin is a substrate for transglutaminases. Biochem Biophys Res Commun. 1988 Nov 30;157(1):115–120. doi: 10.1016/s0006-291x(88)80020-2. [DOI] [PubMed] [Google Scholar]
  59. Selkoe D. J., Abraham C., Ihara Y. Brain transglutaminase: in vitro crosslinking of human neurofilament proteins into insoluble polymers. Proc Natl Acad Sci U S A. 1982 Oct;79(19):6070–6074. doi: 10.1073/pnas.79.19.6070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Sener A., Dunlop M. E., Gomis R., Mathias P. C., Malaisse-Lagae F., Malaisse W. J. Role of transglutaminase in insulin release. Study with glycine and sarcosine methylesters. Endocrinology. 1985 Jul;117(1):237–242. doi: 10.1210/endo-117-1-237. [DOI] [PubMed] [Google Scholar]
  61. Slife C. W., Dorsett M. D., Tillotson M. L. Subcellular location and identification of a large molecular weight substrate for the liver plasma membrane transglutaminase. J Biol Chem. 1986 Mar 5;261(7):3451–3456. [PubMed] [Google Scholar]
  62. Ta B. M., Gallagher G. T., Chakravarty R., Rice R. H. Keratinocyte transglutaminase in human skin and oral mucosa: cytoplasmic localization and uncoupling of differentiation markers. J Cell Sci. 1990 Apr;95(Pt 4):631–638. doi: 10.1242/jcs.95.4.631. [DOI] [PubMed] [Google Scholar]
  63. Takahashi N., Takahashi Y., Putnam F. W. Primary structure of blood coagulation factor XIIIa (fibrinoligase, transglutaminase) from human placenta. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8019–8023. doi: 10.1073/pnas.83.21.8019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Thacher S. M., Rice R. H. Keratinocyte-specific transglutaminase of cultured human epidermal cells: relation to cross-linked envelope formation and terminal differentiation. Cell. 1985 Mar;40(3):685–695. doi: 10.1016/0092-8674(85)90217-x. [DOI] [PubMed] [Google Scholar]
  65. Thomázy V., Fésüs L. Differential expression of tissue transglutaminase in human cells. An immunohistochemical study. Cell Tissue Res. 1989 Jan;255(1):215–224. doi: 10.1007/BF00229084. [DOI] [PubMed] [Google Scholar]
  66. Turner P. M., Lorand L. Complexation of fibronectin with tissue transglutaminase. Biochemistry. 1989 Jan 24;28(2):628–635. doi: 10.1021/bi00428a032. [DOI] [PubMed] [Google Scholar]
  67. Tyrrell D. J., Sale W. S., Slife C. W. Fibronectin is a component of the sodium dodecyl sulfate-insoluble transglutaminase substrate. J Biol Chem. 1988 Jun 15;263(17):8464–8469. [PubMed] [Google Scholar]
  68. Upchurch H. F., Conway E., Patterson M. K., Jr, Birckbichler P. J., Maxwell M. D. Cellular transglutaminase has affinity for extracellular matrix. In Vitro Cell Dev Biol. 1987 Nov;23(11):795–800. doi: 10.1007/BF02623682. [DOI] [PubMed] [Google Scholar]
  69. Upchurch H. F., Conway E., Patterson M. K., Jr, Maxwell M. D. Localization of cellular transglutaminase on the extracellular matrix after wounding: characteristics of the matrix bound enzyme. J Cell Physiol. 1991 Dec;149(3):375–382. doi: 10.1002/jcp.1041490304. [DOI] [PubMed] [Google Scholar]
  70. Williams-Ashman H. G. Transglutaminases and the clotting of mammalian seminal fluids. Mol Cell Biochem. 1984;58(1-2):51–61. doi: 10.1007/BF00240604. [DOI] [PubMed] [Google Scholar]

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