Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Jan;84(1):161–165. doi: 10.1073/pnas.84.1.161

Cloned murine T200 (Ly-5) cDNA reveals multiple transcripts within B- and T-lymphocyte lineages.

W C Raschke
PMCID: PMC304162  PMID: 2948186

Abstract

The murine T200 family of cell-surface glycoproteins is expressed on hematopoietic lineage cell types in a developmentally regulated manner with different lymphocyte subpopulations expressing cross-reactive but structurally distinct forms. To investigate the differences between these forms and the regulation of these differences, murine T200 cDNA clones were isolated by using a probe obtained by subtractive hybridization. This procedure made use of a T200+ L-cell transfectant and the parent Ltk- cell line. The 1.9-kilobase (kb) cDNA clone was sequenced and found to contain the coding region of the COOH-terminal 331 amino acids and 0.9 kb of 3' untranslated region. Where the sequence overlapped with the rat sequence, 80-90% homology was observed. RNA blot analysis revealed that B-lineage cell lines express either a 5.6-kb or a 6.5-kb mRNA correlating to the size difference of the T200 glycoprotein synthesized. Similarly, in the T-cell lineage a helper T-cell clone and a cytotoxic T-cell clone express T200 transcripts of 5.6 kb and 5.9 kb, respectively, which correlate with the distinct sizes of their T200 glycoproteins. A T200-negative mutant of a T-cell line was found to express full-length T200 mRNA, although at a diminished level.

Full text

PDF
161

Images in this article

Selected References

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

  1. Dale R. M., McClure B. A., Houchins J. P. A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. Plasmid. 1985 Jan;13(1):31–40. doi: 10.1016/0147-619x(85)90053-8. [DOI] [PubMed] [Google Scholar]
  2. Hyman R., Trowbridge I. Two complementation classes of T200 (Ly-5) glycoprotein-negative mutants. Immunogenetics. 1981 Mar 1;12(5-6):511–523. doi: 10.1007/BF01561692. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Lizardi P. M., Engelberg A. Rapid isolation of RNA using proteinase K and sodium perchlorate. Anal Biochem. 1979 Sep 15;98(1):116–122. doi: 10.1016/0003-2697(79)90714-0. [DOI] [PubMed] [Google Scholar]
  5. Omary M. B., Trowbridge I. S., Scheid M. P. T200 cell surface glycoprotein of the mouse. Polymorphism defined by the Ly-5 system of alloantigens. J Exp Med. 1980 May 1;151(5):1311–1316. doi: 10.1084/jem.151.5.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Paige C. J., Kincade P. W., Ralph P. Murine B cell leukemia line with inducible surface immunoglobulin expression. J Immunol. 1978 Aug;121(2):641–647. [PubMed] [Google Scholar]
  7. Raschke W. C., Hyman R. Stable expression of the mouse lymphocyte T200 antigen in L-cells after transfection with lymphoma DNA. Mol Immunol. 1985 Oct;22(10):1137–1143. doi: 10.1016/0161-5890(85)90001-x. [DOI] [PubMed] [Google Scholar]
  8. Raschke W. C. Plasmacytomas, lymphomas and hybridomas: their contribution to immunology and molecular biology. Biochim Biophys Acta. 1980 Mar 12;605(1):113–145. doi: 10.1016/0304-419x(80)90023-2. [DOI] [PubMed] [Google Scholar]
  9. Raschke W. C. Transformation by Abelson murine leukemia virus: properties of the transformed cells. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1187–1194. doi: 10.1101/sqb.1980.044.01.128. [DOI] [PubMed] [Google Scholar]
  10. Shen F. W., Saga Y., Litman G., Freeman G., Tung J. S., Cantor H., Boyse E. A. Cloning of Ly-5 cDNA. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7360–7363. doi: 10.1073/pnas.82.21.7360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Trowbridge I. S. Interspecies spleen-myeloma hybrid producing monoclonal antibodies against mouse lymphocyte surface glycoprotein, T200. J Exp Med. 1978 Jul 1;148(1):313–323. doi: 10.1084/jem.148.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Trowbridge I. S., Mazauskas C. Immunological properties of murine thymus-dependent lymphocyte surface glycoproteins. Eur J Immunol. 1976 Aug;6(8):557–562. doi: 10.1002/eji.1830060806. [DOI] [PubMed] [Google Scholar]
  15. Tung J. S., Deere M. C., Boyse E. A. Evidence that Ly-5 product of T and B cells differ in protein structure. Immunogenetics. 1984;19(2):149–154. doi: 10.1007/BF00387858. [DOI] [PubMed] [Google Scholar]
  16. Tung J. S., Scheid M. P., Palladino M. A. Different forms of Ly-5 within the T-cell lineage. Immunogenetics. 1983;17(6):649–654. doi: 10.1007/BF00366132. [DOI] [PubMed] [Google Scholar]
  17. Wall R., Lippman S., Toth K., Fedoroff N. A general method for the large-scale isolation of polysomes and messenger RNA applied to MOPC 21 mouse myeloma tumors. Anal Biochem. 1977 Sep;82(1):115–129. doi: 10.1016/0003-2697(77)90140-3. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES