Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Nov 1;103(5):1671–1677. doi: 10.1083/jcb.103.5.1671

Transformed human cells release different fibronectin variants than do normal cells

PMCID: PMC2114372  PMID: 3023390

Abstract

Fibronectin molecules are dimers composed of subunits whose primary structures may differ. This is due to alternative splicing in at least two regions (ED and IIICS) of the pre-mRNA. Using two monoclonal antibodies specific for two different epitopes of domain 5 (high affinity for heparin), we have quantitatively analyzed the expression of the IIICS sequence in human fibronectins from different sources. The results demonstrated that the percentage of fibronectin subunits containing the IIICS is higher in fibronectins from tumor-derived or simian virus 40-transformed human cells than in fibronectins from human plasma or normal human fibroblasts. Furthermore, we observed that 45- 65% of fibronectin subunits from transformed cells or normal embryonic fibroblasts are sialylated on the heparin-binding domain 5, whereas this occurs in only 24-28% of fibronectin subunits from normal adult fibroblasts. On the contrary, no sialylation was observed on domain 5 in fibronectin from human plasma.

Full Text

The Full Text of this article is available as a PDF (982.7 KB).

Selected References

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

  1. Alitalo K., Vaheri A. Pericellular matrix in malignant transformation. Adv Cancer Res. 1982;37:111–158. doi: 10.1016/s0065-230x(08)60883-0. [DOI] [PubMed] [Google Scholar]
  2. Garcia-Pardo A., Pearlstein E., Frangione B. Primary structure of human plasma fibronectin. Characterization of a 31,000-dalton fragment from the COOH-terminal region containing a free sulfhydryl group and a fibrin-binding site. J Biol Chem. 1985 Aug 25;260(18):10320–10325. [PubMed] [Google Scholar]
  3. Hynes R. O., Yamada K. M. Fibronectins: multifunctional modular glycoproteins. J Cell Biol. 1982 Nov;95(2 Pt 1):369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hynes R. Molecular biology of fibronectin. Annu Rev Cell Biol. 1985;1:67–90. doi: 10.1146/annurev.cb.01.110185.000435. [DOI] [PubMed] [Google Scholar]
  5. Koch G. A., Schoen R. C., Klebe R. J., Shows T. B. Assignment of a fibronection gene to human chromosome 2 using monoclonal antibodies. Exp Cell Res. 1982 Oct;141(2):293–302. doi: 10.1016/0014-4827(82)90217-8. [DOI] [PubMed] [Google Scholar]
  6. Kornblihtt A. R., Umezawa K., Vibe-Pedersen K., Baralle F. E. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 1985 Jul;4(7):1755–1759. doi: 10.1002/j.1460-2075.1985.tb03847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kornblihtt A. R., Vibe-Pedersen K., Baralle F. E. Human fibronectin: cell specific alternative mRNA splicing generates polypeptide chains differing in the number of internal repeats. Nucleic Acids Res. 1984 Jul 25;12(14):5853–5868. doi: 10.1093/nar/12.14.5853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kornblihtt A. R., Vibe-Pedersen K., Baralle F. E. Human fibronectin: molecular cloning evidence for two mRNA species differing by an internal segment coding for a structural domain. EMBO J. 1984 Jan;3(1):221–226. doi: 10.1002/j.1460-2075.1984.tb01787.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kornblihtt A. R., Vibe-Pedersen K., Baralle F. E. Isolation and characterization of cDNA clones for human and bovine fibronectins. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3218–3222. doi: 10.1073/pnas.80.11.3218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Matsuura H., Hakomori S. The oncofetal domain of fibronectin defined by monoclonal antibody FDC-6: its presence in fibronectins from fetal and tumor tissues and its absence in those from normal adult tissues and plasma. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6517–6521. doi: 10.1073/pnas.82.19.6517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Petersen T. E., Thøgersen H. C., Skorstengaard K., Vibe-Pedersen K., Sahl P., Sottrup-Jensen L., Magnusson S. Partial primary structure of bovine plasma fibronectin: three types of internal homology. Proc Natl Acad Sci U S A. 1983 Jan;80(1):137–141. doi: 10.1073/pnas.80.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ruoslahti E., Engvall E., Hayman E. G. Fibronectin: current concepts of its structure and functions. Coll Relat Res. 1981;1(1):95–128. doi: 10.1016/s0174-173x(80)80011-2. [DOI] [PubMed] [Google Scholar]
  15. Schwarzbauer J. E., Paul J. I., Hynes R. O. On the origin of species of fibronectin. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1424–1428. doi: 10.1073/pnas.82.5.1424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schwarzbauer J. E., Tamkun J. W., Lemischka I. R., Hynes R. O. Three different fibronectin mRNAs arise by alternative splicing within the coding region. Cell. 1983 Dec;35(2 Pt 1):421–431. doi: 10.1016/0092-8674(83)90175-7. [DOI] [PubMed] [Google Scholar]
  17. Sekiguchi K., Hakomori S. Domain structure of human plasma fibronectin. Differences and similarities between human and hamster fibronectins. J Biol Chem. 1983 Mar 25;258(6):3967–3973. [PubMed] [Google Scholar]
  18. Sekiguchi K., Siri A., Zardi L., Hakomori S. Differences in domain structure between human fibronectins isolated from plasma and from culture supernatants of normal and transformed fibroblasts. Studies with domain-specific antibodies. J Biol Chem. 1985 Apr 25;260(8):5105–5114. [PubMed] [Google Scholar]
  19. Sekiguchi K., Siri A., Zardi L., Hakomori S. Differences in domain structure between pericellular matrix and plasma fibronectins as revealed by domain-specific antibodies combined with limited proteolysis and S-cyanylation: a preliminary note. Biochem Biophys Res Commun. 1983 Oct 31;116(2):534–540. doi: 10.1016/0006-291x(83)90556-9. [DOI] [PubMed] [Google Scholar]
  20. Tamkun J. W., Schwarzbauer J. E., Hynes R. O. A single rat fibronectin gene generates three different mRNAs by alternative splicing of a complex exon. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5140–5144. doi: 10.1073/pnas.81.16.5140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zardi L., Carnemolla B., Balza E., Borsi L., Castellani P., Rocco M., Siri A. Elution of fibronectin proteolytic fragments from a hydroxyapatite chromatography column. A simple procedure for the purification of fibronectin domains. Eur J Biochem. 1985 Feb 1;146(3):571–579. doi: 10.1111/j.1432-1033.1985.tb08690.x. [DOI] [PubMed] [Google Scholar]
  23. Zardi L., Carnemolla B., Siri A., Santi L., Accolla R. S. Somatic cell hybrids producing antibodies specific to human fibronectin. Int J Cancer. 1980 Mar 15;25(3):325–329. doi: 10.1002/ijc.2910250304. [DOI] [PubMed] [Google Scholar]
  24. Zardi L., Cianfriglia M., Balza E., Carnemolla B., Siri A., Croce C. M. Species-specific monoclonal antibodies in the assignment of the gene for human fibronectin to chromosome 2. EMBO J. 1982;1(8):929–933. doi: 10.1002/j.1460-2075.1982.tb01273.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zardi L., Siri A., Carnemolla B., Cosulich E., Viale G., Santi L. A simplified procedure for the preparation of antibodies to serum fibronectin. J Immunol Methods. 1980;34(2):155–165. doi: 10.1016/0022-1759(80)90169-6. [DOI] [PubMed] [Google Scholar]
  26. Zardi L., Siri A., Carnemolla B., Santi L., Gardner W. D., Hoch S. O. Fibronectin: a chromatin-associated protein? Cell. 1979 Nov;18(3):649–657. doi: 10.1016/0092-8674(79)90120-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES