Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Aug 2;130(4):1005–1014. doi: 10.1083/jcb.130.4.1005

Large and small splice variants of collagen XII: differential expression and ligand binding

PMCID: PMC2199960  PMID: 7642694

Abstract

Collagen XII has a short collagenous tail and a very large, three-armed NC3 domains consisting primarily of fibronectin type III repeats. Differential splicing within this domain gives rise to a large (320 kD) and a small (220 kD) subunit; the large but not the small can carry glycosaminoglycan. To investigate whether collagen XII variants have distinct expression patterns and functions, we generated antibody and cDNA probes specific for the alternatively spliced domain. We report here that the large variant has a more restricted expression in embryonic tissue than the small. For example, whereas the small variant is widespread in the dermis, the large is limited to the base of feather buds. Distinct proportions of mRNA for the two variants were detected depending on the tissue. Monoclonal antibodies allowed us to separate collagen XII variants, and to show that homo- and heterotrimers exist. Collagen XII variants differ in ligand binding. Small subunits interact weakly with heparin via their COOH-terminal domain. Large subunits have additional, stronger heparin-binding site(s) in their NH2-terminal extra domain. In vivo, both large and small collagen XII are associated with interstitial collagen. Here we show biochemically and ultrastructurally that collagen XII can be incorporated into collagen I fibrils when it is present during, but not after, fibril formation. Removal of the collagenous domain of collagen XII reduces its coprecipitation with collagen I. Our results indicate that collagen XII is specifically associated with fibrillar collagen, and that the large variant has binding sites for extracellular ligands not present in the small variant.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

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

  1. Barkalow F. J., Schwarzbauer J. E. Localization of the major heparin-binding site in fibronectin. J Biol Chem. 1991 Apr 25;266(12):7812–7818. [PubMed] [Google Scholar]
  2. Birk D. E., Fitch J. M., Babiarz J. P., Doane K. J., Linsenmayer T. F. Collagen fibrillogenesis in vitro: interaction of types I and V collagen regulates fibril diameter. J Cell Sci. 1990 Apr;95(Pt 4):649–657. doi: 10.1242/jcs.95.4.649. [DOI] [PubMed] [Google Scholar]
  3. Birk D. E., Zycband E. I., Winkelmann D. A., Trelstad R. L. Collagen fibrillogenesis in situ: fibril segments are intermediates in matrix assembly. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4549–4553. doi: 10.1073/pnas.86.12.4549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown J. C., Mann K., Wiedemann H., Timpl R. Structure and binding properties of collagen type XIV isolated from human placenta. J Cell Biol. 1993 Jan;120(2):557–567. doi: 10.1083/jcb.120.2.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brubacher D., Wehrle-Haller B., Chiquet M. Chick laminin: isolation by monoclonal antibodies and differential distribution of variants in the embryo. Exp Cell Res. 1991 Dec;197(2):290–299. doi: 10.1016/0014-4827(91)90435-w. [DOI] [PubMed] [Google Scholar]
  6. Cardin A. D., Weintraub H. J. Molecular modeling of protein-glycosaminoglycan interactions. Arteriosclerosis. 1989 Jan-Feb;9(1):21–32. doi: 10.1161/01.atv.9.1.21. [DOI] [PubMed] [Google Scholar]
  7. Castagnola P., Tavella S., Gerecke D. R., Dublet B., Gordon M. K., Seyer J., Cancedda R., van der Rest M., Olsen B. R. Tissue-specific expression of type XIV collagen--a member of the FACIT class of collagens. Eur J Cell Biol. 1992 Dec;59(2):340–347. [PubMed] [Google Scholar]
  8. Chiquet-Ehrismann R. Anti-adhesive molecules of the extracellular matrix. Curr Opin Cell Biol. 1991 Oct;3(5):800–804. doi: 10.1016/0955-0674(91)90053-2. [DOI] [PubMed] [Google Scholar]
  9. Chiquet-Ehrismann R., Tannheimer M., Koch M., Brunner A., Spring J., Martin D., Baumgartner S., Chiquet M. Tenascin-C expression by fibroblasts is elevated in stressed collagen gels. J Cell Biol. 1994 Dec;127(6 Pt 2):2093–2101. doi: 10.1083/jcb.127.6.2093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chiquet M., Vrucinić-Filipi N., Schenk S., Beck K., Chiquet-Ehrismann R. Isolation of chick tenascin variants and fragments. A C-terminal heparin-binding fragment produced by cleavage of the extra domain from the largest subunit splicing variant. Eur J Biochem. 1991 Jul 15;199(2):379–388. doi: 10.1111/j.1432-1033.1991.tb16134.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Cox K. H., DeLeon D. V., Angerer L. M., Angerer R. C. Detection of mrnas in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev Biol. 1984 Feb;101(2):485–502. doi: 10.1016/0012-1606(84)90162-3. [DOI] [PubMed] [Google Scholar]
  13. Dublet B., Oh S., Sugrue S. P., Gordon M. K., Gerecke D. R., Olsen B. R., van der Rest M. The structure of avian type XII collagen. Alpha 1 (XII) chains contain 190-kDa non-triple helical amino-terminal domains and form homotrimeric molecules. J Biol Chem. 1989 Aug 5;264(22):13150–13156. [PubMed] [Google Scholar]
  14. Engel J. Common structural motifs in proteins of the extracellular matrix. Curr Opin Cell Biol. 1991 Oct;3(5):779–785. doi: 10.1016/0955-0674(91)90050-9. [DOI] [PubMed] [Google Scholar]
  15. Fischer D., Chiquet-Ehrismann R., Bernasconi C., Chiquet M. A single heparin binding region within the fibrinogen-like domain is functional in chick tenascin-C. J Biol Chem. 1995 Feb 17;270(7):3378–3384. doi: 10.1074/jbc.270.7.3378. [DOI] [PubMed] [Google Scholar]
  16. Fleischmajer R., Fisher L. W., MacDonald E. D., Jacobs L., Jr, Perlish J. S., Termine J. D. Decorin interacts with fibrillar collagen of embryonic and adult human skin. J Struct Biol. 1991 Feb;106(1):82–90. doi: 10.1016/1047-8477(91)90065-5. [DOI] [PubMed] [Google Scholar]
  17. Font B., Aubert-Foucher E., Goldschmidt D., Eichenberger D., van der Rest M. Binding of collagen XIV with the dermatan sulfate side chain of decorin. J Biol Chem. 1993 Nov 25;268(33):25015–25018. [PubMed] [Google Scholar]
  18. Fässler R., Schnegelsberg P. N., Dausman J., Shinya T., Muragaki Y., McCarthy M. T., Olsen B. R., Jaenisch R. Mice lacking alpha 1 (IX) collagen develop noninflammatory degenerative joint disease. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5070–5074. doi: 10.1073/pnas.91.11.5070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gerecke D. R., Foley J. W., Castagnola P., Gennari M., Dublet B., Cancedda R., Linsenmayer T. F., van der Rest M., Olsen B. R., Gordon M. K. Type XIV collagen is encoded by alternative transcripts with distinct 5' regions and is a multidomain protein with homologies to von Willebrand's factor, fibronectin, and other matrix proteins. J Biol Chem. 1993 Jun 5;268(16):12177–12184. [PubMed] [Google Scholar]
  20. Gordon M. K., Gerecke D. R., Olsen B. R. Type XII collagen: distinct extracellular matrix component discovered by cDNA cloning. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6040–6044. doi: 10.1073/pnas.84.17.6040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Griffiths G., McDowall A., Back R., Dubochet J. On the preparation of cryosections for immunocytochemistry. J Ultrastruct Res. 1984 Oct;89(1):65–78. doi: 10.1016/s0022-5320(84)80024-6. [DOI] [PubMed] [Google Scholar]
  22. Jiang T. X., Chuong C. M. Mechanism of skin morphogenesis. I. Analyses with antibodies to adhesion molecules tenascin, N-CAM, and integrin. Dev Biol. 1992 Mar;150(1):82–98. doi: 10.1016/0012-1606(92)90009-6. [DOI] [PubMed] [Google Scholar]
  23. Keene D. R., Lunstrum G. P., Morris N. P., Stoddard D. W., Burgeson R. E. Two type XII-like collagens localize to the surface of banded collagen fibrils. J Cell Biol. 1991 May;113(4):971–978. doi: 10.1083/jcb.113.4.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koch M., Bernasconi C., Chiquet M. A major oligomeric fibroblast proteoglycan identified as a novel large form of type-XII collagen. Eur J Biochem. 1992 Aug 1;207(3):847–856. doi: 10.1111/j.1432-1033.1992.tb17116.x. [DOI] [PubMed] [Google Scholar]
  25. Lunstrum G. P., McDonough A. M., Marinkovich M. P., Keene D. R., Morris N. P., Burgeson R. E. Identification and partial purification of a large, variant form of type XII collagen. J Biol Chem. 1992 Oct 5;267(28):20087–20092. [PubMed] [Google Scholar]
  26. Lunstrum G. P., Morris N. P., McDonough A. M., Keene D. R., Burgeson R. E. Identification and partial characterization of two type XII-like collagen molecules. J Cell Biol. 1991 May;113(4):963–969. doi: 10.1083/jcb.113.4.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mayne R., Brewton R. G. New members of the collagen superfamily. Curr Opin Cell Biol. 1993 Oct;5(5):883–890. doi: 10.1016/0955-0674(93)90039-s. [DOI] [PubMed] [Google Scholar]
  28. Ninomiya Y., Olsen B. R. Synthesis and characterization of cDNA encoding a cartilage-specific short collagen. Proc Natl Acad Sci U S A. 1984 May;81(10):3014–3018. doi: 10.1073/pnas.81.10.3014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Oh S. P., Griffith C. M., Hay E. D., Olsen B. R. Tissue-specific expression of type XII collagen during mouse embryonic development. Dev Dyn. 1993 Jan;196(1):37–46. doi: 10.1002/aja.1001960105. [DOI] [PubMed] [Google Scholar]
  30. Paulsson M., Aumailley M., Deutzmann R., Timpl R., Beck K., Engel J. Laminin-nidogen complex. Extraction with chelating agents and structural characterization. Eur J Biochem. 1987 Jul 1;166(1):11–19. doi: 10.1111/j.1432-1033.1987.tb13476.x. [DOI] [PubMed] [Google Scholar]
  31. Salmivirta M., Elenius K., Vainio S., Hofer U., Chiquet-Ehrismann R., Thesleff I., Jalkanen M. Syndecan from embryonic tooth mesenchyme binds tenascin. J Biol Chem. 1991 Apr 25;266(12):7733–7739. [PubMed] [Google Scholar]
  32. Shaw L. M., Olsen B. R. FACIT collagens: diverse molecular bridges in extracellular matrices. Trends Biochem Sci. 1991 May;16(5):191–194. doi: 10.1016/0968-0004(91)90074-6. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Trueb J., Trueb B. The two splice variants of collagen XII share a common 5' end. Biochim Biophys Acta. 1992 Nov 15;1171(1):97–98. doi: 10.1016/0167-4781(92)90145-p. [DOI] [PubMed] [Google Scholar]
  35. Vaughan L., Mendler M., Huber S., Bruckner P., Winterhalter K. H., Irwin M. I., Mayne R. D-periodic distribution of collagen type IX along cartilage fibrils. J Cell Biol. 1988 Mar;106(3):991–997. doi: 10.1083/jcb.106.3.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Watt S. L., Lunstrum G. P., McDonough A. M., Keene D. R., Burgeson R. E., Morris N. P. Characterization of collagen types XII and XIV from fetal bovine cartilage. J Biol Chem. 1992 Oct 5;267(28):20093–20099. [PubMed] [Google Scholar]
  37. Wu J. J., Woods P. E., Eyre D. R. Identification of cross-linking sites in bovine cartilage type IX collagen reveals an antiparallel type II-type IX molecular relationship and type IX to type IX bonding. J Biol Chem. 1992 Nov 15;267(32):23007–23014. [PubMed] [Google Scholar]
  38. Wälchli C., Koch M., Chiquet M., Odermatt B. F., Trueb B. Tissue-specific expression of the fibril-associated collagens XII and XIV. J Cell Sci. 1994 Feb;107(Pt 2):669–681. doi: 10.1242/jcs.107.2.669. [DOI] [PubMed] [Google Scholar]
  39. Wälchli C., Trueb J., Kessler B., Winterhalter K. H., Trueb B. Complete primary structure of chicken collagen XIV. Eur J Biochem. 1993 Mar 1;212(2):483–490. doi: 10.1111/j.1432-1033.1993.tb17685.x. [DOI] [PubMed] [Google Scholar]
  40. Yamagata M., Yamada K. M., Yamada S. S., Shinomura T., Tanaka H., Nishida Y., Obara M., Kimata K. The complete primary structure of type XII collagen shows a chimeric molecule with reiterated fibronectin type III motifs, von Willebrand factor A motifs, a domain homologous to a noncollagenous region of type IX collagen, and short collagenous domains with an Arg-Gly-Asp site. J Cell Biol. 1991 Oct;115(1):209–221. doi: 10.1083/jcb.115.1.209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yurchenco P. D., Sung U., Ward M. D., Yamada Y., O'Rear J. J. Recombinant laminin G domain mediates myoblast adhesion and heparin binding. J Biol Chem. 1993 Apr 15;268(11):8356–8365. [PubMed] [Google Scholar]
  42. van der Rest M., Dublet B., Champliaud M. F. Fibril-associated collagens. Biomaterials. 1990 Jul;11:28–31. [PubMed] [Google Scholar]

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

RESOURCES