Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 May 1;98(5):1662–1671. doi: 10.1083/jcb.98.5.1662

Changes in cell shape correlate with collagenase gene expression in rabbit synovial fibroblasts

PMCID: PMC2113188  PMID: 6327718

Abstract

Induction of the neutral proteinase, collagenase, is a marker for a specific switch in gene expression observed in rabbit synovial fibroblasts. A variety of agents, including 12-O-tetradecanoylphorbol- 13-acetate, cytochalasins B and D, trypsin, chymotrypsin, poly(2- hydroxyethylmethacrylate), and trifluoperazine induced this change in gene expression. Induction of collagenase by these agents was always correlated with a marked alteration in cell morphology, although the cells remained adherent to the culture dishes. The amount of collagenase induced was positively correlated with the degree of shape change produced by a given concentration and, to some extent, with the duration of treatment. Altered cell morphology was required only during the first few hours of treatment with inducing agents; after this time collagenase synthesis continued for up to 6 d even when agents were removed and normal flattened cell morphology was regained. All agents that altered cell morphology also produced a characteristic switch in protein secretion phenotype, characterized by the induction of procollagenase (Mr 53,000 and 57,000) and a neutral metalloproteinase (Mr 51,000), which accounted for approximately 25% and 15% of the protein secreted, respectively. Secretion of another neutral proteinase, plasminogen activator, did not correlate with increased collagenase secretion. In contrast, synthesis and secretion of a number of other polypeptides, including the extracellular matrix proteins, collagen and fibronectin, were concomitantly decreased. That changes in cell shape correlated with a program of gene expression manifested by both degradation and synthesis of extracellular macromolecules may have broad implications in development, repair, and pathologic conditions.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aggeler J., Frisch S. M., Werb Z. Collagenase is a major gene product of induced rabbit synovial fibroblasts. J Cell Biol. 1984 May;98(5):1656–1661. doi: 10.1083/jcb.98.5.1656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aggeler J., Kapp L. N., Tseng S. C., Werb Z. Regulation of protein secretion in Chinese hamster ovary cells by cell cycle position and cell density. Plasminogen activator, procollagen fibronectin. Exp Cell Res. 1982 Jun;139(2):275–283. doi: 10.1016/0014-4827(82)90252-x. [DOI] [PubMed] [Google Scholar]
  3. Aggeler J., Risch J., Werb Z. Expression of the catalytic activity of plasminogen activator under physiologic conditions. Biochim Biophys Acta. 1981 Jun 11;675(1):62–68. doi: 10.1016/0304-4165(81)90069-6. [DOI] [PubMed] [Google Scholar]
  4. Allan M., Harrison P. Co-expression of differentiation markers in hybrids between Friend cells and lymphoid cells and the influence of the cell shape. Cell. 1980 Feb;19(2):437–447. doi: 10.1016/0092-8674(80)90518-8. [DOI] [PubMed] [Google Scholar]
  5. Benya P. D., Shaffer J. D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 1982 Aug;30(1):215–224. doi: 10.1016/0092-8674(82)90027-7. [DOI] [PubMed] [Google Scholar]
  6. Bissell M. J., Hall H. G., Parry G. How does the extracellular matrix direct gene expression? J Theor Biol. 1982 Nov 7;99(1):31–68. doi: 10.1016/0022-5193(82)90388-5. [DOI] [PubMed] [Google Scholar]
  7. Brinckerhoff C. E., Gross R. H., Nagase H., Sheldon L., Jackson R. C., Harris E. D., Jr Increased level of translatable collagenase messenger ribonucleic acid in rabbit synovial fibroblasts treated with phorbol myristate acetate or crystals of monosodium urate monohydrate. Biochemistry. 1982 May 25;21(11):2674–2679. doi: 10.1021/bi00540a015. [DOI] [PubMed] [Google Scholar]
  8. Brinckerhoff C. E., McMillan R. M., Fahey J. V., Harris E. D., Jr Collagenase production by synovial fibroblasts treated with phorbol myristate acetate. Arthritis Rheum. 1979 Oct;22(10):1109–1116. doi: 10.1002/art.1780221010. [DOI] [PubMed] [Google Scholar]
  9. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  10. Cheung W. Y. Calmodulin plays a pivotal role in cellular regulation. Science. 1980 Jan 4;207(4426):19–27. doi: 10.1126/science.6243188. [DOI] [PubMed] [Google Scholar]
  11. Chinkers M., McKanna J. A., Cohen S. Rapid rounding of human epidermoid carcinoma cells A-431 induced by epidermal growth factor. J Cell Biol. 1981 Feb;88(2):422–429. doi: 10.1083/jcb.88.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cohen S., Ushiro H., Stoscheck C., Chinkers M. A native 170,000 epidermal growth factor receptor-kinase complex from shed plasma membrane vesicles. J Biol Chem. 1982 Feb 10;257(3):1523–1531. [PubMed] [Google Scholar]
  13. Dodd N. J., Schor S. L., Rushton G. The effects of a collagenous extracellular matrix on fibroblast membrane organization. An ESR spin label study. Exp Cell Res. 1982 Oct;141(2):421–431. doi: 10.1016/0014-4827(82)90230-0. [DOI] [PubMed] [Google Scholar]
  14. Emerman J. T., Burwen S. J., Pitelka D. R. Substrate properties influencing ultrastructural differentiation of mammary epithelial cells in culture. Tissue Cell. 1979;11(1):109–119. doi: 10.1016/0040-8166(79)90011-9. [DOI] [PubMed] [Google Scholar]
  15. Emerman J. T., Enami J., Pitelka D. R., Nandi S. Hormonal effects on intracellular and secreted casein in cultures of mouse mammary epithelial cells on floating collagen membranes. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4466–4470. doi: 10.1073/pnas.74.10.4466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Farmer S. R., Ben-Ze'av A., Benecke B. J., Penman S. Altered translatability of messenger RNA from suspended anchorage-dependent fibroblasts: reversal upon cell attachment to a surface. Cell. 1978 Oct;15(2):627–637. doi: 10.1016/0092-8674(78)90031-4. [DOI] [PubMed] [Google Scholar]
  17. Folkman J., Moscona A. Role of cell shape in growth control. Nature. 1978 Jun 1;273(5661):345–349. doi: 10.1038/273345a0. [DOI] [PubMed] [Google Scholar]
  18. Gordon S., Werb Z. Secretion of macrophage neutral proteinase is enhanced by colchicine. Proc Natl Acad Sci U S A. 1976 Mar;73(3):872–876. doi: 10.1073/pnas.73.3.872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harris E. D., Jr, Krane S. M. Effects of colchicine on collagenase in cultures of rheumatoid synovium. Arthritis Rheum. 1971 Nov-Dec;14(6):669–684. doi: 10.1002/art.1780140602. [DOI] [PubMed] [Google Scholar]
  20. Harris E. D., Jr, Reynolds J. J., Werb Z. Cytochalasin B increases collagenase production by cells in vitro. Nature. 1975 Sep 18;257(5523):243–244. doi: 10.1038/257243a0. [DOI] [PubMed] [Google Scholar]
  21. Johnson-Muller B., Gross J. Regulation of corneal collagenase production: epithelial-stromal cell interactions. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4417–4421. doi: 10.1073/pnas.75.9.4417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Juva K., Prockop D. J. Modified procedure for the assay of H-3-or C-14-labeled hydroxyproline. Anal Biochem. 1966 Apr;15(1):77–83. doi: 10.1016/0003-2697(66)90249-1. [DOI] [PubMed] [Google Scholar]
  23. Kao K. J., Sommer J. R., Pizzo S. V. Modulation of platelet shape and membrane receptor binding by Ca2+-calmodulin complex. Nature. 1981 Jul 2;292(5818):82–84. doi: 10.1038/292082a0. [DOI] [PubMed] [Google Scholar]
  24. Kenigsberg R. L., Côté A., Trifaró J. M. Trifluoperazine, a calmodulin inhibitor, blocks secretion in cultured chromaffin cells at a step distal from calcium entry. Neuroscience. 1982;7(9):2277–2286. doi: 10.1016/0306-4522(82)90138-5. [DOI] [PubMed] [Google Scholar]
  25. Kraft A. S., Anderson W. B. Phorbol esters increase the amount of Ca2+, phospholipid-dependent protein kinase associated with plasma membrane. Nature. 1983 Feb 17;301(5901):621–623. doi: 10.1038/301621a0. [DOI] [PubMed] [Google Scholar]
  26. Lee L. S., Weinstein I. B. Epidermal growth factor, like phorbol esters, induces plasminogen activator in HeLa cells. Nature. 1978 Aug 17;274(5672):696–697. doi: 10.1038/274696a0. [DOI] [PubMed] [Google Scholar]
  27. Levin R. M., Weiss B. Specificity of the binding of trifluoperazine to the calcium-dependent activator of phosphodiesterase and to a series of other calcium-binding proteins. Biochim Biophys Acta. 1978 May 3;540(2):197–204. doi: 10.1016/0304-4165(78)90132-0. [DOI] [PubMed] [Google Scholar]
  28. Liebermann D., Hoffman-Liebermann B., Sachs L. Regulation of gene expression by tumor promoters. II. Control of cell shape and developmental programs for macrophages and granulocytes in human myeloid leukemic cells. Int J Cancer. 1981 Sep 15;28(3):285–291. doi: 10.1002/ijc.2910280306. [DOI] [PubMed] [Google Scholar]
  29. MacLean-Fletcher S., Pollard T. D. Mechanism of action of cytochalasin B on actin. Cell. 1980 Jun;20(2):329–341. doi: 10.1016/0092-8674(80)90619-4. [DOI] [PubMed] [Google Scholar]
  30. Mizel S. B., Dayer J. M., Krane S. M., Mergenhagen S. E. Stimulation of rheumatoid synovial cell collagenase and prostaglandin production by partially purified lymphocyte-activating factor (interleukin 1). Proc Natl Acad Sci U S A. 1981 Apr;78(4):2474–2477. doi: 10.1073/pnas.78.4.2474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Moscatelli D., Jaffe E., Rifkin D. B. Tetradecanoyl phorbol acetate stimulates latent collagenase production by cultured human endothelial cells. Cell. 1980 Jun;20(2):343–351. doi: 10.1016/0092-8674(80)90620-0. [DOI] [PubMed] [Google Scholar]
  32. Nelson G. A., Andrews M. L., Karnovsky M. J. Control of erythrocyte shape by calmodulin. J Cell Biol. 1983 Mar;96(3):730–735. doi: 10.1083/jcb.96.3.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Penman S., Fulton A., Capco D., Ben Ze'ev A., Wittelsberger S., Tse C. F. Cytoplasmic and nuclear architecture in cells and tissue: form, functions, and mode of assembly. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):1013–1028. doi: 10.1101/sqb.1982.046.01.094. [DOI] [PubMed] [Google Scholar]
  35. Quigley J. P. Phorbol ester-induced morphological changes in transformed chick fibroblasts: evidence for direct catalytic involvement of plasminogen activator. Cell. 1979 May;17(1):131–141. doi: 10.1016/0092-8674(79)90301-5. [DOI] [PubMed] [Google Scholar]
  36. Reaven E. P., Axline S. G. Subplasmalemmal microfilaments and microtubules in resting and phagocytizing cultivated macrophages. J Cell Biol. 1973 Oct;59(1):12–27. doi: 10.1083/jcb.59.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Revel J. P., Hoch P., Ho D. Adhesion of culture cells to their substratum. Exp Cell Res. 1974 Mar 15;84(1):207–218. doi: 10.1016/0014-4827(74)90398-x. [DOI] [PubMed] [Google Scholar]
  38. Rifkin D. B., Crowe R. M., Pollack R. Tumor promoters induce changes in the chick embryo fibroblast cytoskeleton. Cell. 1979 Oct;18(2):361–368. doi: 10.1016/0092-8674(79)90055-2. [DOI] [PubMed] [Google Scholar]
  39. Spiegelman B. M., Green H. Control of specific protein biosynthesis during the adipose conversion of 3T3 cells. J Biol Chem. 1980 Sep 25;255(18):8811–8818. [PubMed] [Google Scholar]
  40. Spruill W. A., Zysk J. R., Tres L. L., Kierszenbaum A. L. Calcium/calmodulin-dependent phosphorylation of vimentin in rat sertoli cells. Proc Natl Acad Sci U S A. 1983 Feb;80(3):760–764. doi: 10.1073/pnas.80.3.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
  42. Tsukita S., Tsukita S., Ishikawa H., Kurokawa M., Morimoto K., Sobue K., Kakiuchi S. Binding sites of calmodulin and actin on the brain spectrin, calspectin. J Cell Biol. 1983 Aug;97(2):574–578. doi: 10.1083/jcb.97.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Unkeless J. C., Tobia A., Ossowski L., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):85–111. doi: 10.1084/jem.137.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Werb Z., Aggeler J. Proteases induce secretion of collagenase and plasminogen activator by fibroblasts. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1839–1843. doi: 10.1073/pnas.75.4.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Werb Z., Mainardi C. L., Vater C. A., Harris E. D., Jr Endogenous activiation of latent collagenase by rheumatoid synovial cells. Evidence for a role of plasminogen activator. N Engl J Med. 1977 May 5;296(18):1017–1023. doi: 10.1056/NEJM197705052961801. [DOI] [PubMed] [Google Scholar]
  46. Werb Z., Reynolds J. J. Stimulation by endocytosis of the secretion of collagenase and neutral proteinase from rabbit synovial fibroblasts. J Exp Med. 1974 Dec 1;140(6):1482–1497. doi: 10.1084/jem.140.6.1482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Woolley D. E., Brinckerhoff C. E., Mainardi C. L., Vater C. A., Evanson J. M., Harris E. D., Jr Collagenase production by rheumatoid synovial cells: morphological and immunohistochemical studies of the dendritic cell. Ann Rheum Dis. 1979 Jun;38(3):262–270. doi: 10.1136/ard.38.3.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Woolley D. E., Harris E. D., Jr, Mainardi C. L., Brinckerhoff C. E. Collagenase immunolocalization in cultures of rheumatoid synovial cells. Science. 1978 May 19;200(4343):773–775. doi: 10.1126/science.205952. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES