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. 1993 Jun 1;121(5):1165–1172. doi: 10.1083/jcb.121.5.1165

Fibronectin binding site in type I collagen regulates fibronectin fibril formation

PMCID: PMC2119696  PMID: 8501121

Abstract

Mov13 fibroblasts, which do not express endogenous alpha 1(I) collagen chains due to a retroviral insertion, were used to study the role of type I collagen in the process of fibronectin fibrillogenesis. While Mov13 cells produced a sparse matrix containing short fibronectin fibrils, transfection with a wild type pro alpha 1(I) collagen gene resulted in the production of an extensive matrix containing fibronectin fibrils of normal length. To study the amino acids involved in the fibronectin-collagen interaction, mutations were introduced into the known fibronectin binding region of the pro alpha 1(I) collagen gene. Substitution of Gln and Ala at positions 774 and 777 of the alpha 1(I) chain for Pro resulted in the formation of short fibronectin fibrils similar to what was observed in untransfected Mov13 cells. Type I collagen carrying these substitutions bound weakly to fibronectin- sepharose and could be eluted off with 1 M urea. The effect of this mutation on fibronectin fibrillogenesis could be rescued by adding either type I collagen or a peptide fragment (CB.7) which contained the wild type fibronectin binding region of the alpha 1(I) chain to the cell culture. These results suggest that fibronectin fibrillogenesis in tissue culture is dependent on type I collagen synthesis, and define an important role for the fibronectin binding site in this process.

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Selected References

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  1. Allen-Hoffmann B. L., Mosher D. F. Matrix assembly sites for exogenous fibronectin are decreased on human fibroblasts after treatment with agents which increase intracellular cAMP. J Biol Chem. 1987 Oct 15;262(29):14361–14365. [PubMed] [Google Scholar]
  2. Bateman J. F., Mascara T., Chan D., Cole W. G. Abnormal type I collagen metabolism by cultured fibroblasts in lethal perinatal osteogenesis imperfecta. Biochem J. 1984 Jan 1;217(1):103–115. doi: 10.1042/bj2170103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benecky M. J., Kolvenbach C. G., Wine R. W., DiOrio J. P., Mosesson M. W. Human plasma fibronectin structure probed by steady-state fluorescence polarization: evidence for a rigid oblate structure. Biochemistry. 1990 Mar 27;29(12):3082–3091. doi: 10.1021/bi00464a027. [DOI] [PubMed] [Google Scholar]
  4. Chen L. B., Murray A., Segal R. A., Bushnell A., Walsh M. L. Studies on intercellular LETS glycoprotein matrices. Cell. 1978 Jun;14(2):377–391. doi: 10.1016/0092-8674(78)90123-x. [DOI] [PubMed] [Google Scholar]
  5. Dessau W., Adelmann B. C., Timpl R. Identification of the sites in collagen alpha-chains that bind serum anti-gelatin factor (cold-insoluble globulin). Biochem J. 1978 Jan 1;169(1):55–59. doi: 10.1042/bj1690055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dzamba B. J., Peters D. M. Arrangement of cellular fibronectin in noncollagenous fibrils in human fibroblast cultures. J Cell Sci. 1991 Nov;100(Pt 3):605–612. doi: 10.1242/jcs.100.3.605. [DOI] [PubMed] [Google Scholar]
  7. Engvall E., Ruoslahti E., Miller E. J. Affinity of fibronectin to collagens of different genetic types and to fibrinogen. J Exp Med. 1978 Jun 1;147(6):1584–1595. doi: 10.1084/jem.147.6.1584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fleischmajer R., Timpl R. Ultrastructural localization of fibronectin to different anatomic structures of human skin. J Histochem Cytochem. 1984 Mar;32(3):315–321. doi: 10.1177/32.3.6693760. [DOI] [PubMed] [Google Scholar]
  9. Fogerty F. J., Mosher D. F. Mechanisms for organization of fibronectin matrix. Cell Differ Dev. 1990 Dec 2;32(3):439–450. doi: 10.1016/0922-3371(90)90061-z. [DOI] [PubMed] [Google Scholar]
  10. Furcht L. T., Smith D., Wendelschafer-Crabb G., Mosher D. F., Foidart J. M. Fibronectin presence in native collagen fibrils of human fibroblasts: immunoperoxidase and immunoferritin localization. J Histochem Cytochem. 1980 Dec;28(12):1319–1333. doi: 10.1177/28.12.7014712. [DOI] [PubMed] [Google Scholar]
  11. Furcht L. T., Wendelschafer-Crabb G., Mosher D. F., Foidart J. M. An axial periodic fibrillar arrangement of antigenic determinants for fibronectin and procollagen on ascorbate treated human fibroblasts. J Supramol Struct. 1980;13(1):15–33. doi: 10.1002/jss.400130103. [DOI] [PubMed] [Google Scholar]
  12. Ingham K. C., Brew S. A., Isaacs B. S. Interaction of fibronectin and its gelatin-binding domains with fluorescent-labeled chains of type I collagen. J Biol Chem. 1988 Apr 5;263(10):4624–4628. [PubMed] [Google Scholar]
  13. Ingham K. C., Brew S. A., Migliorini M. M. Further localization of the gelatin-binding determinants within fibronectin. Active fragments devoid of type II homologous repeat modules. J Biol Chem. 1989 Oct 15;264(29):16977–16980. [PubMed] [Google Scholar]
  14. Irish P. S., Hasty D. L. Immunocytochemical localization of fibronectin in human fibroblast cultures using a cell surface replica technique. J Histochem Cytochem. 1983 Jan;31(1):69–77. doi: 10.1177/31.1.6833740. [DOI] [PubMed] [Google Scholar]
  15. Jilek F., Hörmann H. Cold-insoluble globulin (fibronectin), IV[1-35 affinity to soluble collagen of various types. Hoppe Seylers Z Physiol Chem. 1978 Feb;359(2):247–250. doi: 10.1515/bchm.1978.359.1.247. [DOI] [PubMed] [Google Scholar]
  16. Keski-Oja J., Todaro G. J., Vaheri A. Thrombin affects fibronectin and procollagen in the pericellular matrix of cultured human fibroblasts. Biochim Biophys Acta. 1981 Mar 18;673(3):323–331. doi: 10.1016/0304-4165(81)90463-3. [DOI] [PubMed] [Google Scholar]
  17. Kleinman H. K., McGoodwin E. B. Localization of the cell attachment region in types I and II collagens. Biochem Biophys Res Commun. 1976 Sep 20;72(2):426–432. doi: 10.1016/s0006-291x(76)80060-5. [DOI] [PubMed] [Google Scholar]
  18. Kleinman H. K., McGoodwin E. B., Martin G. R., Klebe R. J., Fietzek P. P., Woolley D. E. Localization of the binding site for cell attachment in the alpha1(I) chain of collagen. J Biol Chem. 1978 Aug 25;253(16):5642–5646. [PubMed] [Google Scholar]
  19. Koteliansky V. E., Glukhova M. A., Bejanian M. V., Smirnov V. N., Filimonov V. V., Zalite O. M., Venyaminov SYu A study of the structure of fibronectin. Eur J Biochem. 1981 Oct;119(3):619–624. doi: 10.1111/j.1432-1033.1981.tb05652.x. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Little C. D., Chen W. T. Masking of extracellular collagen and the co-distribution of collagen and fibronectin during matrix formation by cultured embryonic fibroblasts. J Cell Sci. 1982 Jun;55:35–50. doi: 10.1242/jcs.55.1.35. [DOI] [PubMed] [Google Scholar]
  22. Löhler J., Timpl R., Jaenisch R. Embryonic lethal mutation in mouse collagen I gene causes rupture of blood vessels and is associated with erythropoietic and mesenchymal cell death. Cell. 1984 Sep;38(2):597–607. doi: 10.1016/0092-8674(84)90514-2. [DOI] [PubMed] [Google Scholar]
  23. McDonald J. A. Extracellular matrix assembly. Annu Rev Cell Biol. 1988;4:183–207. doi: 10.1146/annurev.cb.04.110188.001151. [DOI] [PubMed] [Google Scholar]
  24. McDonald J. A., Quade B. J., Broekelmann T. J., LaChance R., Forsman K., Hasegawa E., Akiyama S. Fibronectin's cell-adhesive domain and an amino-terminal matrix assembly domain participate in its assembly into fibroblast pericellular matrix. J Biol Chem. 1987 Mar 5;262(7):2957–2967. [PubMed] [Google Scholar]
  25. McKeown-Longo P. J., Mosher D. F. Interaction of the 70,000-mol-wt amino-terminal fragment of fibronectin with the matrix-assembly receptor of fibroblasts. J Cell Biol. 1985 Feb;100(2):364–374. doi: 10.1083/jcb.100.2.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morla A., Ruoslahti E. A fibronectin self-assembly site involved in fibronectin matrix assembly: reconstruction in a synthetic peptide. J Cell Biol. 1992 Jul;118(2):421–429. doi: 10.1083/jcb.118.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Owens R. J., Baralle F. E. Mapping the collagen-binding site of human fibronectin by expression in Escherichia coli. EMBO J. 1986 Nov;5(11):2825–2830. doi: 10.1002/j.1460-2075.1986.tb04575.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Peters D. M., Portz L. M., Fullenwider J., Mosher D. F. Co-assembly of plasma and cellular fibronectins into fibrils in human fibroblast cultures. J Cell Biol. 1990 Jul;111(1):249–256. doi: 10.1083/jcb.111.1.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Quade B. J., McDonald J. A. Fibronectin's amino-terminal matrix assembly site is located within the 29-kDa amino-terminal domain containing five type I repeats. J Biol Chem. 1988 Dec 25;263(36):19602–19609. [PubMed] [Google Scholar]
  30. Schnieke A., Dziadek M., Bateman J., Mascara T., Harbers K., Gelinas R., Jaenisch R. Introduction of the human pro alpha 1(I) collagen gene into pro alpha 1(I)-deficient Mov-13 mouse cells leads to formation of functional mouse-human hybrid type I collagen. Proc Natl Acad Sci U S A. 1987 Feb;84(3):764–768. doi: 10.1073/pnas.84.3.764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schnieke A., Harbers K., Jaenisch R. Embryonic lethal mutation in mice induced by retrovirus insertion into the alpha 1(I) collagen gene. 1983 Jul 28-Aug 3Nature. 304(5924):315–320. doi: 10.1038/304315a0. [DOI] [PubMed] [Google Scholar]
  32. Schwarzbauer J. E. Identification of the fibronectin sequences required for assembly of a fibrillar matrix. J Cell Biol. 1991 Jun;113(6):1463–1473. doi: 10.1083/jcb.113.6.1463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sottile J., Schwarzbauer J., Selegue J., Mosher D. F. Five type I modules of fibronectin form a functional unit that binds to fibroblasts and Staphylococcus aureus. J Biol Chem. 1991 Jul 15;266(20):12840–12843. [PubMed] [Google Scholar]
  34. Vaheri A., Kurkinen M., Lehto V. P., Linder E., Timpl R. Codistribution of pericellular matrix proteins in cultured fibroblasts and loss in transformation: fibronectin and procollagen. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4944–4948. doi: 10.1073/pnas.75.10.4944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Williams E. C., Janmey P. A., Ferry J. D., Mosher D. F. Conformational states of fibronectin. Effects of pH, ionic strength, and collagen binding. J Biol Chem. 1982 Dec 25;257(24):14973–14978. [PubMed] [Google Scholar]
  36. Wu H., Byrne M. H., Stacey A., Goldring M. B., Birkhead J. R., Jaenisch R., Krane S. M. Generation of collagenase-resistant collagen by site-directed mutagenesis of murine pro alpha 1(I) collagen gene. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5888–5892. doi: 10.1073/pnas.87.15.5888. [DOI] [PMC free article] [PubMed] [Google Scholar]

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