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. 1987 Nov 1;166(5):1548–1566. doi: 10.1084/jem.166.5.1548

Differential usage of three exons generates at least five different mRNAs encoding human leukocyte common antigens

PMCID: PMC2189669  PMID: 2824653

Abstract

Leukocyte common antigens (LCAs, also known as T200 and CD 45) are integral membrane proteins expressed exclusively on hematopoietic cells. These molecules exhibit varying molecular masses and epitopes when expressed in different cell types. To determine the genetic bases for the generation of this diversity, three classes of human LCA cDNA clones that are different near their 5' ends have been isolated. These differences arose as a result of differential usage of three exons as determined from an analysis of a genomic DNA clone. Furthermore, Northern blot analysis with LCA exon-specific probes demonstrates the existence of at least two more LCA mRNA forms that are generated by differential splicing. A comparison of the human and mouse LCA protein sequences revealed a marked difference only in the extracellular domain.

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Selected References

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  1. Barclay A. N., Jackson D. I., Willis A. C., Williams A. F. Lymphocyte specific heterogeneity in the rat leucocyte common antigen (T200) is due to differences in polypeptide sequences near the NH2-terminus. EMBO J. 1987 May;6(5):1259–1264. doi: 10.1002/j.1460-2075.1987.tb02362.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bargmann C. I., Hung M. C., Weinberg R. A. The neu oncogene encodes an epidermal growth factor receptor-related protein. Nature. 1986 Jan 16;319(6050):226–230. doi: 10.1038/319226a0. [DOI] [PubMed] [Google Scholar]
  3. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  4. Berger A. E., Davis J. E., Cresswell P. A human leukocyte antigen identified by a monoclonal antibody. Hum Immunol. 1981 Nov;3(3):231–245. doi: 10.1016/0198-8859(81)90020-3. [DOI] [PubMed] [Google Scholar]
  5. Breitbart R. E., Nguyen H. T., Medford R. M., Destree A. T., Mahdavi V., Nadal-Ginard B. Intricate combinatorial patterns of exon splicing generate multiple regulated troponin T isoforms from a single gene. Cell. 1985 May;41(1):67–82. doi: 10.1016/0092-8674(85)90062-5. [DOI] [PubMed] [Google Scholar]
  6. Childs R. A., Dalchau R., Scudder P., Hounsell E. F., Fabre J. W., Feizi T. Evidence for the occurrence of O-glycosidically linked oligosaccharides of poly-N-acetyllactosamine type on the human leucocyte common antigen. Biochem Biophys Res Commun. 1983 Jan 27;110(2):424–431. doi: 10.1016/0006-291x(83)91166-x. [DOI] [PubMed] [Google Scholar]
  7. Coussens L., Van Beveren C., Smith D., Chen E., Mitchell R. L., Isacke C. M., Verma I. M., Ullrich A. Structural alteration of viral homologue of receptor proto-oncogene fms at carboxyl terminus. Nature. 1986 Mar 20;320(6059):277–280. doi: 10.1038/320277a0. [DOI] [PubMed] [Google Scholar]
  8. Dalchau R., Fabre J. W. Identification with a monoclonal antibody of a predominantly B lymphocyte-specific determinant of the human leukocyte common antigen. Evidence for structural and possible functional diversity of the human leukocyte common molecule. J Exp Med. 1981 Apr 1;153(4):753–765. doi: 10.1084/jem.153.4.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hemperly J. J., Murray B. A., Edelman G. M., Cunningham B. A. Sequence of a cDNA clone encoding the polysialic acid-rich and cytoplasmic domains of the neural cell adhesion molecule N-CAM. Proc Natl Acad Sci U S A. 1986 May;83(9):3037–3041. doi: 10.1073/pnas.83.9.3037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Houssaint E., Tobin S., Cihak J., Lösch U. A chicken leukocyte common antigen: biochemical characterization and ontogenetic study. Eur J Immunol. 1987 Feb;17(2):287–290. doi: 10.1002/eji.1830170221. [DOI] [PubMed] [Google Scholar]
  11. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  12. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  13. Lawn R. M., Fritsch E. F., Parker R. C., Blake G., Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. doi: 10.1016/0092-8674(78)90043-0. [DOI] [PubMed] [Google Scholar]
  14. Ledbetter J. A., Rose L. M., Spooner C. E., Beatty P. G., Martin P. J., Clark E. A. Antibodies to common leukocyte antigen p220 influence human T cell proliferation by modifying IL 2 receptor expression. J Immunol. 1985 Sep;135(3):1819–1825. [PubMed] [Google Scholar]
  15. Leff S. E., Rosenfeld M. G., Evans R. M. Complex transcriptional units: diversity in gene expression by alternative RNA processing. Annu Rev Biochem. 1986;55:1091–1117. doi: 10.1146/annurev.bi.55.070186.005303. [DOI] [PubMed] [Google Scholar]
  16. Lefrancois L., Thomas M. L., Bevan M. J., Trowbridge I. S. Different classes of T lymphocytes have different mRNAs for the leukocyte-common antigen, T200. J Exp Med. 1986 May 1;163(5):1337–1342. doi: 10.1084/jem.163.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lefrançois L., Bevan M. J. Novel antigenic determinants of the T200 glycoprotein expressed preferentially by activated cytotoxic T lymphocytes. J Immunol. 1985 Jul;135(1):374–383. [PubMed] [Google Scholar]
  18. Littman D. R. The structure of the CD4 and CD8 genes. Annu Rev Immunol. 1987;5:561–584. doi: 10.1146/annurev.iy.05.040187.003021. [DOI] [PubMed] [Google Scholar]
  19. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  20. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Morimoto C., Letvin N. L., Distaso J. A., Aldrich W. R., Schlossman S. F. The isolation and characterization of the human suppressor inducer T cell subset. J Immunol. 1985 Mar;134(3):1508–1515. [PubMed] [Google Scholar]
  22. Morishima Y., Ogata S., Collins N. H., Dupont B., Lloyd K. O. Carbohydrate differences in human high molecular weight antigens of B- and T-cell lines. Immunogenetics. 1982;15(5):529–535. doi: 10.1007/BF00345912. [DOI] [PubMed] [Google Scholar]
  23. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nakauchi H., Nolan G. P., Hsu C., Huang H. S., Kavathas P., Herzenberg L. A. Molecular cloning of Lyt-2, a membrane glycoprotein marking a subset of mouse T lymphocytes: molecular homology to its human counterpart, Leu-2/T8, and to immunoglobulin variable regions. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5126–5130. doi: 10.1073/pnas.82.15.5126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nieminen P., Saksela E. NK-9, a distinct sialylated antigen of the T200 family. Eur J Immunol. 1986 May;16(5):513–518. doi: 10.1002/eji.1830160509. [DOI] [PubMed] [Google Scholar]
  26. Omary M. B., Trowbridge I. S., Battifora H. A. Human homologue of murine T200 glycoprotein. J Exp Med. 1980 Oct 1;152(4):842–852. doi: 10.1084/jem.152.4.842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Omary M. B., Trowbridge I. S. Disposition of T200 glycoprotein in the plasma membrane of a murine lymphoma cell line. J Biol Chem. 1980 Feb 25;255(4):1662–1669. [PubMed] [Google Scholar]
  28. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  29. Ralph S. J., Thomas M. L., Morton C. C., Trowbridge I. S. Structural variants of human T200 glycoprotein (leukocyte-common antigen). EMBO J. 1987 May;6(5):1251–1257. doi: 10.1002/j.1460-2075.1987.tb02361.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Raschke W. C. Cloned murine T200 (Ly-5) cDNA reveals multiple transcripts within B- and T-lymphocyte lineages. Proc Natl Acad Sci U S A. 1987 Jan;84(1):161–165. doi: 10.1073/pnas.84.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reinherz E. L., Schlossman S. F. The differentiation and function of human T lymphocytes. Cell. 1980 Apr;19(4):821–827. doi: 10.1016/0092-8674(80)90072-0. [DOI] [PubMed] [Google Scholar]
  32. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  33. Saga Y., Tung J. S., Shen F. W., Boyse E. A. Sequences of Ly-5 cDNA: isoform-related diversity of Ly-5 mRNA. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6940–6944. doi: 10.1073/pnas.83.18.6940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sarmiento M., Loken M. R., Trowbridge I. S., Coffman R. L., Fitch F. W. High molecular weight lymphocyte surface proteins are structurally related and are expressed on different cell populations at different times during lymphocyte maturation and differentiation. J Immunol. 1982 Apr;128(4):1676–1684. [PubMed] [Google Scholar]
  35. Shackelford D. A., Trowbridge I. S. Identification of lymphocyte integral membrane proteins as substrates for protein kinase C. Phosphorylation of the interleukin-2 receptor, class I HLA antigens, and T200 glycoprotein. J Biol Chem. 1986 Jun 25;261(18):8334–8341. [PubMed] [Google Scholar]
  36. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  37. Spickett G. P., Brandon M. R., Mason D. W., Williams A. F., Woollett G. R. MRC OX-22, a monoclonal antibody that labels a new subset of T lymphocytes and reacts with the high molecular weight form of the leukocyte-common antigen. J Exp Med. 1983 Sep 1;158(3):795–810. doi: 10.1084/jem.158.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Takeuchi T., Rudd C. E., Schlossman S. F., Morimoto C. Induction of suppression following autologous mixed lymphocyte reaction; role of a novel 2H4 antigen. Eur J Immunol. 1987 Jan;17(1):97–103. doi: 10.1002/eji.1830170117. [DOI] [PubMed] [Google Scholar]
  39. Thomas M. L., Barclay A. N., Gagnon J., Williams A. F. Evidence from cDNA clones that the rat leukocyte-common antigen (T200) spans the lipid bilayer and contains a cytoplasmic domain of 80,000 Mr. Cell. 1985 May;41(1):83–93. doi: 10.1016/0092-8674(85)90063-7. [DOI] [PubMed] [Google Scholar]
  40. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  41. Tung J. S., Scheid M. P., Pierotti M. A., Hämmerling U., Boyse E. A. Structural features and selective expression of three Ly-5+ cell-surface molecules. Immunogenetics. 1981;14(1-2):101–106. doi: 10.1007/BF00344303. [DOI] [PubMed] [Google Scholar]
  42. Ullrich A., Coussens L., Hayflick J. S., Dull T. J., Gray A., Tam A. W., Lee J., Yarden Y., Libermann T. A., Schlessinger J. Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells. 1984 May 31-Jun 6Nature. 309(5967):418–425. doi: 10.1038/309418a0. [DOI] [PubMed] [Google Scholar]
  43. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weis J. J., Fearon D. T., Klickstein L. B., Wong W. W., Richards S. A., de Bruyn Kops A., Smith J. A., Weis J. H. Identification of a partial cDNA clone for the C3d/Epstein-Barr virus receptor of human B lymphocytes: homology with the receptor for fragments C3b and C4b of the third and fourth components of complement. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5639–5643. doi: 10.1073/pnas.83.15.5639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Woollett G. R., Barclay A. N., Puklavec M., Williams A. F. Molecular and antigenic heterogeneity of the rat leukocyte-common antigen from thymocytes and T and B lymphocytes. Eur J Immunol. 1985 Feb;15(2):168–173. doi: 10.1002/eji.1830150211. [DOI] [PubMed] [Google Scholar]
  46. Yamamoto T., Ikawa S., Akiyama T., Semba K., Nomura N., Miyajima N., Saito T., Toyoshima K. Similarity of protein encoded by the human c-erb-B-2 gene to epidermal growth factor receptor. Nature. 1986 Jan 16;319(6050):230–234. doi: 10.1038/319230a0. [DOI] [PubMed] [Google Scholar]
  47. Yarden Y., Escobedo J. A., Kuang W. J., Yang-Feng T. L., Daniel T. O., Tremble P. M., Chen E. Y., Ando M. E., Harkins R. N., Francke U. Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986 Sep 18;323(6085):226–232. doi: 10.1038/323226a0. [DOI] [PubMed] [Google Scholar]
  48. Zamoyska R., Vollmer A. C., Sizer K. C., Liaw C. W., Parnes J. R. Two Lyt-2 polypeptides arise from a single gene by alternative splicing patterns of mRNA. Cell. 1985 Nov;43(1):153–163. doi: 10.1016/0092-8674(85)90020-0. [DOI] [PubMed] [Google Scholar]
  49. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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