Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 May 15;300(Pt 1):69–74. doi: 10.1042/bj3000069

A new alternative transcript encodes a 60 kDa truncated form of integrin beta 3.

I Djaffar 1, Y P Chen 1, C Creminon 1, J Maclouf 1, A M Cieutat 1, O Gayet 1, J P Rosa 1
PMCID: PMC1138124  PMID: 8198553

Abstract

A cDNA for integrin beta 3 isolated from a human erythroleukaemia (HEL) cell library contained a 340 bp insert at position 1281. This mRNA, termed beta 3c, results from the use of a cryptic AG donor splice site in intron 8 of the beta 3 gene, and is different from a previously described alternative beta 3 mRNA. The predicted open reading frame of beta 3C stops at a TAG stop codon 69 bp downstream from position 1281. It starts with the signal peptide and the 404 N-terminal extracellular residues of beta 3, encompassing the ligand binding sites, followed by 23 C-terminal intron-derived residues, corresponding to a truncated form of beta 3 lacking the cysteine-rich, transmembrane and cytoplasmic domains. Expression of beta 3C mRNA was demonstrated in human platelets, megakaryocytes, endothelial cells and HEL cells by reverse transcriptase/PCR. The beta 3C transcript was also demonstrated in the mouse, suggesting its conservation through evolution. Finally, a 60 kDa polypeptide corresponding to the beta 3C alternative transcript was demonstrated in platelets by Western blotting using a polyclonal antibody raised against a synthetic peptide designed from the beta 3C intronic sequence. Taken together, these results suggest a biological role for beta 3C, the first alternative transcript showing an altered extracellular domain of a beta integrin.

Full text

PDF
74

Images in this article

71Figure 1
on p.71
72Figure 2
on p.72
72Figure 3
on p.72
72Figure 4
on p.72

Selected References

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

  1. Albelda S. M., Buck C. A. Integrins and other cell adhesion molecules. FASEB J. 1990 Aug;4(11):2868–2880. [PubMed] [Google Scholar]
  2. Altruda F., Cervella P., Tarone G., Botta C., Balzac F., Stefanuto G., Silengo L. A human integrin beta 1 subunit with a unique cytoplasmic domain generated by alternative mRNA processing. Gene. 1990 Nov 15;95(2):261–266. doi: 10.1016/0378-1119(90)90369-3. [DOI] [PubMed] [Google Scholar]
  3. Argraves W. S., Suzuki S., Arai H., Thompson K., Pierschbacher M. D., Ruoslahti E. Amino acid sequence of the human fibronectin receptor. J Cell Biol. 1987 Sep;105(3):1183–1190. doi: 10.1083/jcb.105.3.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Balzac F., Belkin A. M., Koteliansky V. E., Balabanov Y. V., Altruda F., Silengo L., Tarone G. Expression and functional analysis of a cytoplasmic domain variant of the beta 1 integrin subunit. J Cell Biol. 1993 Apr;121(1):171–178. doi: 10.1083/jcb.121.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bray P. F., Leung C. S., Shuman M. A. Human platelets and megakaryocytes contain alternately spliced glycoprotein IIb mRNAs. J Biol Chem. 1990 Jun 15;265(17):9587–9590. [PubMed] [Google Scholar]
  6. Calvete J. J., Mann K., Alvarez M. V., López M. M., González-Rodríguez J. Proteolytic dissection of the isolated platelet fibrinogen receptor, integrin GPIIb/IIIa. Localization of GPIIb and GPIIIa sequences putatively involved in the subunit interface and in intrasubunit and intrachain contacts. Biochem J. 1992 Mar 1;282(Pt 2):523–532. doi: 10.1042/bj2820523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Charo I. F., Nannizzi L., Phillips D. R., Hsu M. A., Scarborough R. M. Inhibition of fibrinogen binding to GP IIb-IIIa by a GP IIIa peptide. J Biol Chem. 1991 Jan 25;266(3):1415–1421. [PubMed] [Google Scholar]
  8. Chen Y. P., Djaffar I., Pidard D., Steiner B., Cieutat A. M., Caen J. P., Rosa J. P. Ser-752-->Pro mutation in the cytoplasmic domain of integrin beta 3 subunit and defective activation of platelet integrin alpha IIb beta 3 (glycoprotein IIb-IIIa) in a variant of Glanzmann thrombasthenia. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10169–10173. doi: 10.1073/pnas.89.21.10169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cieutat A. M., Rosa J. P., Letourneur F., Poncz M., Rifat S. A comparative analysis of cDNA-derived sequences for rat and mouse beta 3 integrins (GPIIIA) with their human counterpart. Biochem Biophys Res Commun. 1993 Jun 15;193(2):771–778. doi: 10.1006/bbrc.1993.1692. [DOI] [PubMed] [Google Scholar]
  10. D'Souza S. E., Ginsberg M. H., Burke T. A., Lam S. C., Plow E. F. Localization of an Arg-Gly-Asp recognition site within an integrin adhesion receptor. Science. 1988 Oct 7;242(4875):91–93. doi: 10.1126/science.3262922. [DOI] [PubMed] [Google Scholar]
  11. D'Souza S. E., Ginsberg M. H., Matsueda G. R., Plow E. F. A discrete sequence in a platelet integrin is involved in ligand recognition. Nature. 1991 Mar 7;350(6313):66–68. doi: 10.1038/350066a0. [DOI] [PubMed] [Google Scholar]
  12. Djaffar I., Vilette D., Bray P. F., Rosa J. P. Quantitative isolation of RNA from human platelets. Thromb Res. 1991 May 1;62(3):127–135. doi: 10.1016/0049-3848(91)90187-2. [DOI] [PubMed] [Google Scholar]
  13. Erle D. J., Rüegg C., Sheppard D., Pytela R. Complete amino acid sequence of an integrin beta subunit (beta 7) identified in leukocytes. J Biol Chem. 1991 Jun 15;266(17):11009–11016. [PubMed] [Google Scholar]
  14. Fitzgerald L. A., Steiner B., Rall S. C., Jr, Lo S. S., Phillips D. R. Protein sequence of endothelial glycoprotein IIIa derived from a cDNA clone. Identity with platelet glycoprotein IIIa and similarity to "integrin". J Biol Chem. 1987 Mar 25;262(9):3936–3939. [PubMed] [Google Scholar]
  15. Hogervorst F., Kuikman I., von dem Borne A. E., Sonnenberg A. Cloning and sequence analysis of beta-4 cDNA: an integrin subunit that contains a unique 118 kd cytoplasmic domain. EMBO J. 1990 Mar;9(3):765–770. doi: 10.1002/j.1460-2075.1990.tb08171.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  17. Kajiji S., Tamura R. N., Quaranta V. A novel integrin (alpha E beta 4) from human epithelial cells suggests a fourth family of integrin adhesion receptors. EMBO J. 1989 Mar;8(3):673–680. doi: 10.1002/j.1460-2075.1989.tb03425.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kishimoto T. K., O'Connor K., Lee A., Roberts T. M., Springer T. A. Cloning of the beta subunit of the leukocyte adhesion proteins: homology to an extracellular matrix receptor defines a novel supergene family. Cell. 1987 Feb 27;48(4):681–690. doi: 10.1016/0092-8674(87)90246-7. [DOI] [PubMed] [Google Scholar]
  19. Languino L. R., Ruoslahti E. An alternative form of the integrin beta 1 subunit with a variant cytoplasmic domain. J Biol Chem. 1992 Apr 5;267(10):7116–7120. [PubMed] [Google Scholar]
  20. McLaughlin L. L., Wei Y. F., Stockmann P. T., Leahy K. M., Needleman P., Grassi J., Pradelles P. Development, validation and application of an enzyme immunoassay (EIA) of atriopeptin. Biochem Biophys Res Commun. 1987 Apr 14;144(1):469–476. doi: 10.1016/s0006-291x(87)80533-8. [DOI] [PubMed] [Google Scholar]
  21. McLean J. W., Vestal D. J., Cheresh D. A., Bodary S. C. cDNA sequence of the human integrin beta 5 subunit. J Biol Chem. 1990 Oct 5;265(28):17126–17131. [PubMed] [Google Scholar]
  22. Moyle M., Napier M. A., McLean J. W. Cloning and expression of a divergent integrin subunit beta 8. J Biol Chem. 1991 Oct 15;266(29):19650–19658. [PubMed] [Google Scholar]
  23. Otey C. A., Pavalko F. M., Burridge K. An interaction between alpha-actinin and the beta 1 integrin subunit in vitro. J Cell Biol. 1990 Aug;111(2):721–729. doi: 10.1083/jcb.111.2.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Phillips D. R., Charo I. F., Scarborough R. M. GPIIb-IIIa: the responsive integrin. Cell. 1991 May 3;65(3):359–362. doi: 10.1016/0092-8674(91)90451-4. [DOI] [PubMed] [Google Scholar]
  25. Ramaswamy H., Hemler M. E. Cloning, primary structure and properties of a novel human integrin beta subunit. EMBO J. 1990 May;9(5):1561–1568. doi: 10.1002/j.1460-2075.1990.tb08275.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosa J. P., Bray P. F., Gayet O., Johnston G. I., Cook R. G., Jackson K. W., Shuman M. A., McEver R. P. Cloning of glycoprotein IIIa cDNA from human erythroleukemia cells and localization of the gene to chromosome 17. Blood. 1988 Aug;72(2):593–600. [PubMed] [Google Scholar]
  27. Rosa J. P., McEver R. P. Processing and assembly of the integrin, glycoprotein IIb-IIIa, in HEL cells. J Biol Chem. 1989 Jul 25;264(21):12596–12603. [PubMed] [Google Scholar]
  28. Ruoslahti E. Integrins. J Clin Invest. 1991 Jan;87(1):1–5. doi: 10.1172/JCI114957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  30. Sheppard D., Rozzo C., Starr L., Quaranta V., Erle D. J., Pytela R. Complete amino acid sequence of a novel integrin beta subunit (beta 6) identified in epithelial cells using the polymerase chain reaction. J Biol Chem. 1990 Jul 15;265(20):11502–11507. [PubMed] [Google Scholar]
  31. Smith J. W., Cheresh D. A. The Arg-Gly-Asp binding domain of the vitronectin receptor. Photoaffinity cross-linking implicates amino acid residues 61-203 of the beta subunit. J Biol Chem. 1988 Dec 15;263(35):18726–18731. [PubMed] [Google Scholar]
  32. Suzuki S., Huang Z. S., Tanihara H. Cloning of an integrin beta subunit exhibiting high homology with integrin beta 3 subunit. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5354–5358. doi: 10.1073/pnas.87.14.5354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Suzuki S., Naitoh Y. Amino acid sequence of a novel integrin beta 4 subunit and primary expression of the mRNA in epithelial cells. EMBO J. 1990 Mar;9(3):757–763. doi: 10.1002/j.1460-2075.1990.tb08170.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tamura R. N., Rozzo C., Starr L., Chambers J., Reichardt L. F., Cooper H. M., Quaranta V. Epithelial integrin alpha 6 beta 4: complete primary structure of alpha 6 and variant forms of beta 4. J Cell Biol. 1990 Oct;111(4):1593–1604. doi: 10.1083/jcb.111.4.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tamura R. N., Rozzo C., Starr L., Chambers J., Reichardt L. F., Cooper H. M., Quaranta V. Epithelial integrin alpha 6 beta 4: complete primary structure of alpha 6 and variant forms of beta 4. J Cell Biol. 1990 Oct;111(4):1593–1604. doi: 10.1083/jcb.111.4.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yuan Q. A., Jiang W. M., Krissansen G. W., Watson J. D. Cloning and sequence analysis of a novel beta 2-related integrin transcript from T lymphocytes: homology of integrin cysteine-rich repeats to domain III of laminin B chains. Int Immunol. 1990;2(11):1097–1108. doi: 10.1093/intimm/2.11.1097. [DOI] [PubMed] [Google Scholar]
  37. Zimrin A. B., Eisman R., Vilaire G., Schwartz E., Bennett J. S., Poncz M. Structure of platelet glycoprotein IIIa. A common subunit for two different membrane receptors. J Clin Invest. 1988 May;81(5):1470–1475. doi: 10.1172/JCI113478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zimrin A. B., Gidwitz S., Lord S., Schwartz E., Bennett J. S., White G. C., 2nd, Poncz M. The genomic organization of platelet glycoprotein IIIa. J Biol Chem. 1990 May 25;265(15):8590–8595. [PubMed] [Google Scholar]
  39. van Kuppevelt T. H., Languino L. R., Gailit J. O., Suzuki S., Ruoslahti E. An alternative cytoplasmic domain of the integrin beta 3 subunit. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5415–5418. doi: 10.1073/pnas.86.14.5415. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES