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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Feb 1;99(3):520–525. doi: 10.1172/JCI119188

Alternative expression of platelet glycoprotein Ib(beta) mRNA from an adjacent 5' gene with an imperfect polyadenylation signal sequence.

B Zieger 1, Y Hashimoto 1, J Ware 1
PMCID: PMC507827  PMID: 9022087

Abstract

Glycoprotein (GP) Ib is a major component of the platelet membrane receptor for von Willebrand factor, designated the GP Ib-IX-V complex. GP Ib is composed of two subunits (GP Ib(alpha) and GP Ib(beta)) each synthesized from separate genes. The 206 amino acid precursor of GP Ib(beta) is synthesized from a 1.0-kb mRNA expressed by megakaryocytes and was originally characterized from cDNA clones of human erythroleukemic (HEL) cell mRNA, a cell line exhibiting megakaryocytic-like properties. The cell line CHRF-288-11 also exhibits megakaryocytic-like properties, but synthesizes two related GP Ib(beta) mRNA species of 3.5 and 1.0 kb. We performed cDNA cloning experiments to identify the origin of the 3.5-kb transcript and determine its relationship to the 1.0-kb GP Ib(beta) mRNA found in megakaryocytes, platelets, and HEL cells. Our cloning experiments demonstrate that the longer transcript results from a nonconsensus polyadenylation recognition sequence, 5'AACAAT3', within a separate gene located upstream to the platelet GP Ib(beta) gene. In the absence of normal polyadenylation the more 5' gene uses the polyadenylation site within its 3' neighbor, the platelet GP Ib(beta) gene. This newly identified 5' gene contains an open reading frame encoding 369 amino acids with a high degree of sequence similarity to an expanding family of GTP-binding proteins.

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Selected References

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

  1. Adams M. D., Kelley J. M., Gocayne J. D., Dubnick M., Polymeropoulos M. H., Xiao H., Merril C. R., Wu A., Olde B., Moreno R. F. Complementary DNA sequencing: expressed sequence tags and human genome project. Science. 1991 Jun 21;252(5013):1651–1656. doi: 10.1126/science.2047873. [DOI] [PubMed] [Google Scholar]
  2. Bourne H. R., Sanders D. A., McCormick F. The GTPase superfamily: conserved structure and molecular mechanism. Nature. 1991 Jan 10;349(6305):117–127. doi: 10.1038/349117a0. [DOI] [PubMed] [Google Scholar]
  3. Budarf M. L., Konkle B. A., Ludlow L. B., Michaud D., Li M., Yamashiro D. J., McDonald-McGinn D., Zackai E. H., Driscoll D. A. Identification of a patient with Bernard-Soulier syndrome and a deletion in the DiGeorge/velo-cardio-facial chromosomal region in 22q11.2. Hum Mol Genet. 1995 Apr;4(4):763–766. doi: 10.1093/hmg/4.4.763. [DOI] [PubMed] [Google Scholar]
  4. Driscoll D. A. Genetic basis of DiGeorge and velocardiofacial syndromes. Curr Opin Pediatr. 1994 Dec;6(6):702–706. doi: 10.1097/00008480-199412000-00016. [DOI] [PubMed] [Google Scholar]
  5. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  6. Fugman D. A., Witte D. P., Jones C. L., Aronow B. J., Lieberman M. A. In vitro establishment and characterization of a human megakaryoblastic cell line. Blood. 1990 Mar 15;75(6):1252–1261. [PubMed] [Google Scholar]
  7. Hartwell L. H., Culotti J., Pringle J. R., Reid B. J. Genetic control of the cell division cycle in yeast. Science. 1974 Jan 11;183(4120):46–51. doi: 10.1126/science.183.4120.46. [DOI] [PubMed] [Google Scholar]
  8. Hartwell L. H. Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis. Exp Cell Res. 1971 Dec;69(2):265–276. doi: 10.1016/0014-4827(71)90223-0. [DOI] [PubMed] [Google Scholar]
  9. Hickey M. J., Deaven L. L., Roth G. J. Human platelet glycoprotein IX. Characterization of cDNA and localization of the gene to chromosome 3. FEBS Lett. 1990 Nov 12;274(1-2):189–192. doi: 10.1016/0014-5793(90)81361-q. [DOI] [PubMed] [Google Scholar]
  10. Kato Kikuya. A Collection of cDNA Clones with Specific Expression Patterns in Mouse Brain. Eur J Neurosci. 1990;2(8):704–711. doi: 10.1111/j.1460-9568.1990.tb00460.x. [DOI] [PubMed] [Google Scholar]
  11. Kelly M. D., Essex D. W., Shapiro S. S., Meloni F. J., Druck T., Huebner K., Konkle B. A. Complementary DNA cloning of the alternatively expressed endothelial cell glycoprotein Ib beta (GPIb beta) and localization of the GPIb beta gene to chromosome 22. J Clin Invest. 1994 Jun;93(6):2417–2424. doi: 10.1172/JCI117249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  15. Lindsay E. A., Goldberg R., Jurecic V., Morrow B., Carlson C., Kucherlapati R. S., Shprintzen R. J., Baldini A. Velo-cardio-facial syndrome: frequency and extent of 22q11 deletions. Am J Med Genet. 1995 Jul 3;57(3):514–522. doi: 10.1002/ajmg.1320570339. [DOI] [PubMed] [Google Scholar]
  16. Lopez J. A., Chung D. W., Fujikawa K., Hagen F. S., Davie E. W., Roth G. J. The alpha and beta chains of human platelet glycoprotein Ib are both transmembrane proteins containing a leucine-rich amino acid sequence. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2135–2139. doi: 10.1073/pnas.85.7.2135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lopez J. A., Chung D. W., Fujikawa K., Hagen F. S., Papayannopoulou T., Roth G. J. Cloning of the alpha chain of human platelet glycoprotein Ib: a transmembrane protein with homology to leucine-rich alpha 2-glycoprotein. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5615–5619. doi: 10.1073/pnas.84.16.5615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. López J. A., Leung B., Reynolds C. C., Li C. Q., Fox J. E. Efficient plasma membrane expression of a functional platelet glycoprotein Ib-IX complex requires the presence of its three subunits. J Biol Chem. 1992 Jun 25;267(18):12851–12859. [PubMed] [Google Scholar]
  19. López J. A. The platelet glycoprotein Ib-IX complex. Blood Coagul Fibrinolysis. 1994 Feb;5(1):97–119. [PubMed] [Google Scholar]
  20. Macejak D. G., Sarnow P. Internal initiation of translation mediated by the 5' leader of a cellular mRNA. Nature. 1991 Sep 5;353(6339):90–94. doi: 10.1038/353090a0. [DOI] [PubMed] [Google Scholar]
  21. Mountford P. S., Smith A. G. Internal ribosome entry sites and dicistronic RNAs in mammalian transgenesis. Trends Genet. 1995 May;11(5):179–184. doi: 10.1016/S0168-9525(00)89040-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Neufeld T. P., Rubin G. M. The Drosophila peanut gene is required for cytokinesis and encodes a protein similar to yeast putative bud neck filament proteins. Cell. 1994 May 6;77(3):371–379. doi: 10.1016/0092-8674(94)90152-x. [DOI] [PubMed] [Google Scholar]
  23. Nottenburg C., Gallatin W. M., St John T. Lymphocyte HEV adhesion variants differ in the expression of multiple gene sequences. Gene. 1990 Nov 15;95(2):279–284. doi: 10.1016/0378-1119(90)90372-x. [DOI] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Saraste M., Sibbald P. R., Wittinghofer A. The P-loop--a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci. 1990 Nov;15(11):430–434. doi: 10.1016/0968-0004(90)90281-f. [DOI] [PubMed] [Google Scholar]
  26. Sheets M. D., Ogg S. C., Wickens M. P. Point mutations in AAUAAA and the poly (A) addition site: effects on the accuracy and efficiency of cleavage and polyadenylation in vitro. Nucleic Acids Res. 1990 Oct 11;18(19):5799–5805. doi: 10.1093/nar/18.19.5799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yagi M., Edelhoff S., Disteche C. M., Roth G. J. Structural characterization and chromosomal location of the gene encoding human platelet glycoprotein Ib beta. J Biol Chem. 1994 Jul 1;269(26):17424–17427. [PubMed] [Google Scholar]

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