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. 1996 Feb 1;313(Pt 3):879–884. doi: 10.1042/bj3130879

Cellular activation of mesangial gelatinase A by cytochalasin D is accompanied by enhanced mRNA expression of both gelatinase A and its membrane-associated gelatinase A activator (MT-MMP).

M Ailenberg 1, M Silverman 1
PMCID: PMC1216993  PMID: 8611170

Abstract

Activation of gelatinase A represents a crucial regulatory step in the control of its enzymic activity. Rat kidney mesangial cells secrete predominantly latent gelatinase A that can be activated following treatment with cytochalasin D. In the present paper we provide new evidence, using reverse transcription-PCR, that treatment of rat mesangial cells with cytochalasin D enhances the steady-state level of mRNA of the membrane-type matrix metalloproteinase (MT-MMP), as well as of gelatinase A, with no change in the level of tissue inhibitor of metalloproteinases-2 (TIMP-2) mRNA. Since the TIMP-2 protein level is reduced in conditioned medium from cytochalasin D-treated cells, the results of the present study are consistent with a model in which the action of cytochalasin D is to cause extracellular gelatinase A and TIMP-2 to be sequestered at the plasma membrane, forming a heterotrimeric complex with MT-MMP. In this manner, TIMP-2 may assume a bifunctional role causing: (i) inhibition of gelatinase A in the extracellular compartment; and (ii) guiding gelatinase A to activation through a membrane association with MT-MMP.

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Selected References

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  1. Ailenberg M., Stetler-Stevenson W. G., Fritz I. B. Secretion of latent type IV procollagenase and active type IV collagenase by testicular cells in culture. Biochem J. 1991 Oct 1;279(Pt 1):75–80. doi: 10.1042/bj2790075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allenberg M., Weinstein T., Li I., Silverman M. Activation of procollagenase IV by cytochalasin D and concanavalin A in cultured rat mesangial cells: linkage to cytoskeletal reorganization. J Am Soc Nephrol. 1994 Apr;4(10):1760–1770. doi: 10.1681/ASN.V4101760. [DOI] [PubMed] [Google Scholar]
  3. Baricos W. H., Shah S. V. Proteolytic enzymes as mediators of glomerular injury. Kidney Int. 1991 Aug;40(2):161–173. doi: 10.1038/ki.1991.196. [DOI] [PubMed] [Google Scholar]
  4. Brown P. D., Kleiner D. E., Unsworth E. J., Stetler-Stevenson W. G. Cellular activation of the 72 kDa type IV procollagenase/TIMP-2 complex. Kidney Int. 1993 Jan;43(1):163–170. doi: 10.1038/ki.1993.27. [DOI] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Davies M., Thomas G. J., Martin J., Lovett D. H. The purification and characterization of a glomerular-basement-membrane-degrading neutral proteinase from rat mesangial cells. Biochem J. 1988 Apr 15;251(2):419–425. doi: 10.1042/bj2510419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emonard H. P., Remacle A. G., Noël A. C., Grimaud J. A., Stetler-Stevenson W. G., Foidart J. M. Tumor cell surface-associated binding site for the M(r) 72,000 type IV collagenase. Cancer Res. 1992 Oct 15;52(20):5845–5848. [PubMed] [Google Scholar]
  8. Goldberg G. I., Marmer B. L., Grant G. A., Eisen A. Z., Wilhelm S., He C. S. Human 72-kilodalton type IV collagenase forms a complex with a tissue inhibitor of metalloproteases designated TIMP-2. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8207–8211. doi: 10.1073/pnas.86.21.8207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heussen C., Dowdle E. B. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem. 1980 Feb;102(1):196–202. doi: 10.1016/0003-2697(80)90338-3. [DOI] [PubMed] [Google Scholar]
  10. Liotta L. A., Tryggvason K., Garbisa S., Hart I., Foltz C. M., Shafie S. Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature. 1980 Mar 6;284(5751):67–68. doi: 10.1038/284067a0. [DOI] [PubMed] [Google Scholar]
  11. Marti H. P., McNeil L., Davies M., Martin J., Lovett D. H. Homology cloning of rat 72 kDa type IV collagenase: cytokine and second-messenger inducibility in glomerular mesangial cells. Biochem J. 1993 Apr 15;291(Pt 2):441–446. doi: 10.1042/bj2910441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Murphy G., Willenbrock F., Ward R. V., Cockett M. I., Eaton D., Docherty A. J. The C-terminal domain of 72 kDa gelatinase A is not required for catalysis, but is essential for membrane activation and modulates interactions with tissue inhibitors of metalloproteinases. Biochem J. 1992 May 1;283(Pt 3):637–641. doi: 10.1042/bj2830637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Okada A., Bellocq J. P., Rouyer N., Chenard M. P., Rio M. C., Chambon P., Basset P. Membrane-type matrix metalloproteinase (MT-MMP) gene is expressed in stromal cells of human colon, breast, and head and neck carcinomas. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2730–2734. doi: 10.1073/pnas.92.7.2730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Overall C. M., Sodek J. Concanavalin A produces a matrix-degradative phenotype in human fibroblasts. Induction and endogenous activation of collagenase, 72-kDa gelatinase, and Pump-1 is accompanied by the suppression of the tissue inhibitor of matrix metalloproteinases. J Biol Chem. 1990 Dec 5;265(34):21141–21151. [PubMed] [Google Scholar]
  15. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sato H., Takino T., Okada Y., Cao J., Shinagawa A., Yamamoto E., Seiki M. A matrix metalloproteinase expressed on the surface of invasive tumour cells. Nature. 1994 Jul 7;370(6484):61–65. doi: 10.1038/370061a0. [DOI] [PubMed] [Google Scholar]
  17. Strongin A. Y., Collier I., Bannikov G., Marmer B. L., Grant G. A., Goldberg G. I. Mechanism of cell surface activation of 72-kDa type IV collagenase. Isolation of the activated form of the membrane metalloprotease. J Biol Chem. 1995 Mar 10;270(10):5331–5338. doi: 10.1074/jbc.270.10.5331. [DOI] [PubMed] [Google Scholar]
  18. Strongin A. Y., Marmer B. L., Grant G. A., Goldberg G. I. Plasma membrane-dependent activation of the 72-kDa type IV collagenase is prevented by complex formation with TIMP-2. J Biol Chem. 1993 Jul 5;268(19):14033–14039. [PubMed] [Google Scholar]
  19. Ward R. V., Atkinson S. J., Slocombe P. M., Docherty A. J., Reynolds J. J., Murphy G. Tissue inhibitor of metalloproteinases-2 inhibits the activation of 72 kDa progelatinase by fibroblast membranes. Biochim Biophys Acta. 1991 Aug 30;1079(2):242–246. doi: 10.1016/0167-4838(91)90132-j. [DOI] [PubMed] [Google Scholar]

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