Abstract
We have examined the role of cell surface glycosaminoglycans in fibronectin-mediated cell adhesion by analyzing the adhesive properties of Chinese hamster ovary cell mutants deficient in glycosaminoglycans. The results of our study suggest that the absence of glycosaminoglycans does not affect the initial attachment and subsequent spreading of these cells on substrata composed of intact fibronectin or a fibronectin fragment containing the primary cell-binding domain. However, in contrast to wild-type cells, the glycosaminoglycan- deficient cells did not attach to substrate composed of a heparin- binding fibronectin fragment. Furthermore, the wild-type but not the glycosaminoglycan-deficient cells formed F-actin-containing stress fibers and focal adhesions on substrata composed of intact fibronectin. We propose, therefore, that cell surface proteoglycan(s) participate in the transmembrane linking of intracellular cytoskeletal components to extracellular matrix components which occurs in focal adhesions.
Full Text
The Full Text of this article is available as a PDF (1.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Couchman J. R., Hök M., Rees D. A., Timpl R. Adhesion, growth, and matrix production by fibroblasts on laminin substrates. J Cell Biol. 1983 Jan;96(1):177–183. doi: 10.1083/jcb.96.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Culp L. A., Laterra J., Lark M. W., Beyth R. J., Tobey S. L. Heparan sulphate proteoglycan as mediator of some adhesive responses and cytoskeletal reorganization of cells on fibronectin matrices: independent versus cooperative functions. Ciba Found Symp. 1986;124:158–183. doi: 10.1002/9780470513385.ch10. [DOI] [PubMed] [Google Scholar]
- Dedhar S., Ruoslahti E., Pierschbacher M. D. A cell surface receptor complex for collagen type I recognizes the Arg-Gly-Asp sequence. J Cell Biol. 1987 Mar;104(3):585–593. doi: 10.1083/jcb.104.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Engvall E., Ruoslahti E. Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int J Cancer. 1977 Jul 15;20(1):1–5. doi: 10.1002/ijc.2910200102. [DOI] [PubMed] [Google Scholar]
- Esko J. D., Elgavish A., Prasthofer T., Taylor W. H., Weinke J. L. Sulfate transport-deficient mutants of Chinese hamster ovary cells. Sulfation of glycosaminoglycans dependent on cysteine. J Biol Chem. 1986 Nov 25;261(33):15725–15733. [PubMed] [Google Scholar]
- Esko J. D., Stewart T. E., Taylor W. H. Animal cell mutants defective in glycosaminoglycan biosynthesis. Proc Natl Acad Sci U S A. 1985 May;82(10):3197–3201. doi: 10.1073/pnas.82.10.3197. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esko J. D., Weinke J. L., Taylor W. H., Ekborg G., Rodén L., Anantharamaiah G., Gawish A. Inhibition of chondroitin and heparan sulfate biosynthesis in Chinese hamster ovary cell mutants defective in galactosyltransferase I. J Biol Chem. 1987 Sep 5;262(25):12189–12195. [PubMed] [Google Scholar]
- Gill P. J., Silbert C. K., Silbert J. E. Effects of heparan sulfate removal on attachment and reattachment of fibroblasts and endothelial cells. Biochemistry. 1986 Jan 28;25(2):405–410. doi: 10.1021/bi00350a020. [DOI] [PubMed] [Google Scholar]
- Grinnell F. Cellular adhesiveness and extracellular substrata. Int Rev Cytol. 1978;53:65–144. doi: 10.1016/s0074-7696(08)62241-x. [DOI] [PubMed] [Google Scholar]
- HAM R. G. CLONAL GROWTH OF MAMMALIAN CELLS IN A CHEMICALLY DEFINED, SYNTHETIC MEDIUM. Proc Natl Acad Sci U S A. 1965 Feb;53:288–293. doi: 10.1073/pnas.53.2.288. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horwitz A., Duggan K., Buck C., Beckerle M. C., Burridge K. Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage. Nature. 1986 Apr 10;320(6062):531–533. doi: 10.1038/320531a0. [DOI] [PubMed] [Google Scholar]
- Hynes R. O. Fibronectins. Sci Am. 1986 Jun;254(6):42–51. doi: 10.1038/scientificamerican0686-42. [DOI] [PubMed] [Google Scholar]
- Hök M., Kjellén L., Johansson S. Cell-surface glycosaminoglycans. Annu Rev Biochem. 1984;53:847–869. doi: 10.1146/annurev.bi.53.070184.004215. [DOI] [PubMed] [Google Scholar]
- Hök M., Woods A., Johansson S., Kjellén L., Couchman J. R. Functions of proteoglycans at the cell surface. Ciba Found Symp. 1986;124:143–157. doi: 10.1002/9780470513385.ch9. [DOI] [PubMed] [Google Scholar]
- Izzard C. S., Radinsky R., Culp L. A. Substratum contacts and cytoskeletal reorganization of BALB/c 3T3 cells on a cell-binding fragment and heparin-binding fragments of plasma fibronectin. Exp Cell Res. 1986 Aug;165(2):320–336. doi: 10.1016/0014-4827(86)90586-0. [DOI] [PubMed] [Google Scholar]
- Johansson S., Kjellén L., Hök M., Timpl R. Substrate adhesion of rat hepatocytes: a comparison of laminin and fibronectin as attachment proteins. J Cell Biol. 1981 Jul;90(1):260–264. doi: 10.1083/jcb.90.1.260. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lark M. W., Culp L. A. Multiple classes of heparan sulfate proteoglycans from fibroblast substratum adhesion sites. Affinity fractionation on columns of platelet factor 4, plasma fibronectin, and octyl-sepharose. J Biol Chem. 1984 Jun 10;259(11):6773–6782. [PubMed] [Google Scholar]
- Miekka S. I., Ingham K. C., Menache D. Rapid methods for isolation of human plasma fibronectin. Thromb Res. 1982 Jul 1;27(1):1–14. doi: 10.1016/0049-3848(82)90272-9. [DOI] [PubMed] [Google Scholar]
- Mosher D. F. Physiology of fibronectin. Annu Rev Med. 1984;35:561–575. doi: 10.1146/annurev.me.35.020184.003021. [DOI] [PubMed] [Google Scholar]
- Pytela R., Pierschbacher M. D., Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. doi: 10.1016/0092-8674(85)90322-8. [DOI] [PubMed] [Google Scholar]
- Rapraeger A., Jalkanen M., Bernfield M. Cell surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells. J Cell Biol. 1986 Dec;103(6 Pt 2):2683–2696. doi: 10.1083/jcb.103.6.2683. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg L. C., Choi H. U., Poole A. R., Lewandowska K., Culp L. A. Biological roles of dermatan sulphate proteoglycans. Ciba Found Symp. 1986;124:47–68. doi: 10.1002/9780470513385.ch4. [DOI] [PubMed] [Google Scholar]
- Ruoslahti E., Pierschbacher M. D. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. doi: 10.1016/0092-8674(86)90259-x. [DOI] [PubMed] [Google Scholar]
- Schwarz M. A., Juliano R. L. Two distinct mechanisms for the interaction of cells with fibronectin substrata. J Cell Physiol. 1985 Jul;124(1):113–119. doi: 10.1002/jcp.1041240118. [DOI] [PubMed] [Google Scholar]
- Shively J. E., Conrad H. E. Formation of anhydrosugars in the chemical depolymerization of heparin. Biochemistry. 1976 Sep 7;15(18):3932–3942. doi: 10.1021/bi00663a005. [DOI] [PubMed] [Google Scholar]
- Singer I. I., Kawka D. W., Scott S., Mumford R. A., Lark M. W. The fibronectin cell attachment sequence Arg-Gly-Asp-Ser promotes focal contact formation during early fibroblast attachment and spreading. J Cell Biol. 1987 Mar;104(3):573–584. doi: 10.1083/jcb.104.3.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Streeter H. B., Rees D. A. Fibroblast adhesion to RGDS shows novel features compared with fibronectin. J Cell Biol. 1987 Jul;105(1):507–515. doi: 10.1083/jcb.105.1.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tamkun J. W., DeSimone D. W., Fonda D., Patel R. S., Buck C., Horwitz A. F., Hynes R. O. Structure of integrin, a glycoprotein involved in the transmembrane linkage between fibronectin and actin. Cell. 1986 Jul 18;46(2):271–282. doi: 10.1016/0092-8674(86)90744-0. [DOI] [PubMed] [Google Scholar]
- Vuento M., Vaheri A. Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions. Biochem J. 1979 Nov 1;183(2):331–337. doi: 10.1042/bj1830331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wasteson A., Uthne K., Westermark B. A novel assay for the biosynthesis of sulphated polysaccharide and its application to studies on the effects of somatomedin on cultured cells. Biochem J. 1973 Dec;136(4):1069–1074. doi: 10.1042/bj1361069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woods A., Couchman J. R., Hök M. Heparan sulfate proteoglycans of rat embryo fibroblasts. A hydrophobic form may link cytoskeleton and matrix components. J Biol Chem. 1985 Sep 5;260(19):10872–10879. [PubMed] [Google Scholar]
- Woods A., Couchman J. R., Johansson S., Hök M. Adhesion and cytoskeletal organisation of fibroblasts in response to fibronectin fragments. EMBO J. 1986 Apr;5(4):665–670. doi: 10.1002/j.1460-2075.1986.tb04265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woods A., Hök M., Kjellén L., Smith C. G., Rees D. A. Relationship of heparan sulfate proteoglycans to the cytoskeleton and extracellular matrix of cultured fibroblasts. J Cell Biol. 1984 Nov;99(5):1743–1753. doi: 10.1083/jcb.99.5.1743. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamada K. M. Cell surface interactions with extracellular materials. Annu Rev Biochem. 1983;52:761–799. doi: 10.1146/annurev.bi.52.070183.003553. [DOI] [PubMed] [Google Scholar]