Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1990 Dec;9(12):3839–3847. doi: 10.1002/j.1460-2075.1990.tb07602.x

Activated T cells transcribe an alternatively spliced mRNA encoding a soluble form of Qa-2 antigen.

N Ulker 1, K D Lewis 1, L E Hood 1, I Stroynowski 1
PMCID: PMC552151  PMID: 2249652

Abstract

Among the best characterized non-classical mouse major histocompatibility antigens are the Qa-2 molecules. These proteins can serve as targets for allogenic cytotoxic T cells and as signal transducing molecules. They are structurally similar to H-2 transplantation antigens in their N-terminal and beta 2-microglobulin binding domains but differ at their C-termini. While the H-2 antigens span the cell membrane, the Qa-2 molecules are attached to the cell surface via phospholipid anchors. The genetic information encoding this attachment is contained in exon 5. In concanavalin A activated splenocytes the expression of membrane bound Qa-2 antigens declines and, simultaneously, soluble forms of Qa-2 molecules are secreted. We demonstrate here that the soluble Qa-2 polypeptides are translated from alternatively spliced mRNAs lacking exon 5, while the membrane forms are encoded by the full-size transcripts. In cultured cells the alternative splicing of the Qa-2 message is induced by T-cell activation splicing of the Qa-2 message is induced by T-cell activation with concanavalin A. The canonical mRNA encoding the membrane form of Qa-2 predominates in unstimulated mouse tissues but the cultured cell lines, like activated T cells, express enhanced levels of the truncated mRNA. In some cell lines almost all Qa-2 transcripts lack exon 5. For example, in L cells, mRNAs encoding soluble Qa-2 molecules are at least 10 times more abundant than Qa-2 transcripts encoding phospholipid anchored antigens. These findings are discussed in terms of potential functions of membrane bound and secreted Qa-2 molecules.

Full text

PDF
3839

Images in this article

3840Fig. 1.
on p.3840
3841Fig. 2.
on p.3841
3841Fig. 3.
on p.3841
3842Fig. 4.
on p.3842
3844Fig. 6.
on p.3844
3844Fig. 7.
on p.3844

Selected References

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

  1. Asarnow D. M., Kuziel W. A., Bonyhadi M., Tigelaar R. E., Tucker P. W., Allison J. P. Limited diversity of gamma delta antigen receptor genes of Thy-1+ dendritic epidermal cells. Cell. 1988 Dec 2;55(5):837–847. doi: 10.1016/0092-8674(88)90139-0. [DOI] [PubMed] [Google Scholar]
  2. Bertoncello I., Bartelmez S. H., Bradley T. R., Hodgson G. S. Increased Qa-m7 antigen expression is characteristic of primitive hemopoietic progenitors in regenerating marrow. J Immunol. 1987 Aug 15;139(4):1096–1103. [PubMed] [Google Scholar]
  3. Birkeland M. L., Johnson P., Trowbridge I. S., Puré E. Changes in CD45 isoform expression accompany antigen-induced murine T-cell activation. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6734–6738. doi: 10.1073/pnas.86.17.6734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bjorkman P. J., Saper M. A., Samraoui B., Bennett W. S., Strominger J. L., Wiley D. C. The foreign antigen binding site and T cell recognition regions of class I histocompatibility antigens. Nature. 1987 Oct 8;329(6139):512–518. doi: 10.1038/329512a0. [DOI] [PubMed] [Google Scholar]
  5. Bluestone J. A., Cron R. Q., Cotterman M., Houlden B. A., Matis L. A. Structure and specificity of T cell receptor gamma/delta on major histocompatibility complex antigen-specific CD3+, CD4-, CD8- T lymphocytes. J Exp Med. 1988 Nov 1;168(5):1899–1916. doi: 10.1084/jem.168.5.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Devlin J. J., Weiss E. H., Paulson M., Flavell R. A. Duplicated gene pairs and alleles of class I genes in the Qa2 region of the murine major histocompatibility complex: a comparison. EMBO J. 1985 Dec 1;4(12):3203–3207. doi: 10.1002/j.1460-2075.1985.tb04066.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Early P., Rogers J., Davis M., Calame K., Bond M., Wall R., Hood L. Two mRNAs can be produced from a single immunoglobulin mu gene by alternative RNA processing pathways. Cell. 1980 Jun;20(2):313–319. doi: 10.1016/0092-8674(80)90617-0. [DOI] [PubMed] [Google Scholar]
  9. Fahrner K., Hogan B. L., Flavell R. A. Transcription of H-2 and Qa genes in embryonic and adult mice. EMBO J. 1987 May;6(5):1265–1271. doi: 10.1002/j.1460-2075.1987.tb02363.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Flaherty L., DiBiase K., Lynes M. A., Seidman J. G., Weinberger O., Rinchik E. M. Characterization of a Q subregion gene in the murine major histocompatibility complex. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1503–1507. doi: 10.1073/pnas.82.5.1503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galli G., Guise J., Tucker P. W., Nevins J. R. Poly(A) site choice rather than splice site choice governs the regulated production of IgM heavy-chain RNAs. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2439–2443. doi: 10.1073/pnas.85.8.2439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  13. Hahn A. B., Soloski M. J. Anti-Qa-2-induced T cell activation. The parameters of activation, the definition of mitogenic and nonmitogenic antibodies, and the differential effects on CD4+ vs CD8+ T cells. J Immunol. 1989 Jul 15;143(2):407–413. [PubMed] [Google Scholar]
  14. Hedley M. L., Drake B. L., Head J. R., Tucker P. W., Forman J. Differential expression of the class I MHC genes in the embryo and placenta during midgestational development in the mouse. J Immunol. 1989 Jun 1;142(11):4046–4053. [PubMed] [Google Scholar]
  15. Ito K., Bonneville M., Takagaki Y., Nakanishi N., Kanagawa O., Krecko E. G., Tonegawa S. Different gamma delta T-cell receptors are expressed on thymocytes at different stages of development. Proc Natl Acad Sci U S A. 1989 Jan;86(2):631–635. doi: 10.1073/pnas.86.2.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Krangel M. S. Secretion of HLA-A and -B antigens via an alternative RNA splicing pathway. J Exp Med. 1986 May 1;163(5):1173–1190. doi: 10.1084/jem.163.5.1173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lalanne J. L., Transy C., Guerin S., Darche S., Meulien P., Kourilsky P. Expression of class I genes in the major histocompatibility complex: identification of eight distinct mRNAs in DBA/2 mouse liver. Cell. 1985 Jun;41(2):469–478. doi: 10.1016/s0092-8674(85)80020-9. [DOI] [PubMed] [Google Scholar]
  18. Lew A. M., Maloy W. L., Coligan J. E. Characteristics of the expression of the murine soluble class I molecule (Q10). J Immunol. 1986 Jan;136(1):254–258. [PubMed] [Google Scholar]
  19. Loveland B., Wang C. R., Yonekawa H., Hermel E., Lindahl K. F. Maternally transmitted histocompatibility antigen of mice: a hydrophobic peptide of a mitochondrially encoded protein. Cell. 1990 Mar 23;60(6):971–980. doi: 10.1016/0092-8674(90)90345-f. [DOI] [PubMed] [Google Scholar]
  20. Matsuura A., Schloss R., Shen F. W., Tung J. S., Hunt S. W., 3rd, Fisher D. A., Hood L. E., Boyse E. A. Expression of the Q8/9d gene by T cells of the mouse. Immunogenetics. 1989;30(3):156–161. doi: 10.1007/BF02421200. [DOI] [PubMed] [Google Scholar]
  21. McCluskey J., Boyd L. F., Maloy W. L., Coligan J. E., Margulies D. H. Alternative processing of H-2Dd pre-mRNAs results in membrane expression of differentially phosphorylated protein products. EMBO J. 1986 Oct;5(10):2477–2483. doi: 10.1002/j.1460-2075.1986.tb04524.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Norment A. M., Lonberg N., Lacy E., Littman D. R. Alternatively spliced mRNA encodes a secreted form of human CD8 alpha. Characterization of the human CD8 alpha gene. J Immunol. 1989 May 1;142(9):3312–3319. [PubMed] [Google Scholar]
  23. Ostrand-Rosenberg S., Nickerson D. A., Clements V. K., Garcia E. P., Lamouse-Smith E., Hood L., Stroynowski I. Embryonal carcinoma cells express Qa and Tla class I genes of the major histocompatibility complex. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5084–5088. doi: 10.1073/pnas.86.13.5084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rabinowitz R., Sharrow S. O., Chatterjee-Das S., Rogers M. J., Sachs D. H. Qa alloantigen expression on functional T lymphocytes from spleen and thymus. Immunogenetics. 1986;24(6):391–401. doi: 10.1007/BF00377958. [DOI] [PubMed] [Google Scholar]
  25. Robinson P. J., Millrain M., Antoniou J., Simpson E., Mellor A. L. A glycophospholipid anchor is required for Qa-2-mediated T cell activation. Nature. 1989 Nov 2;342(6245):85–87. doi: 10.1038/342085a0. [DOI] [PubMed] [Google Scholar]
  26. Rogers J., Early P., Carter C., Calame K., Bond M., Hood L., Wall R. Two mRNAs with different 3' ends encode membrane-bound and secreted forms of immunoglobulin mu chain. Cell. 1980 Jun;20(2):303–312. doi: 10.1016/0092-8674(80)90616-9. [DOI] [PubMed] [Google Scholar]
  27. Rogers M. J., Siwarski D. F., Shacter E., Maloy W. L., Lillehoj E. P., Coligan J. E. Three distinct H-2Ks molecules differing at the carboxy terminus are expressed on a tumor from SJL/J mice. J Immunol. 1986 Nov 1;137(9):3006–3012. [PubMed] [Google Scholar]
  28. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Sharrow S. O., Flaherty L., Sachs D. H. Serologic cross-reactivity between Class I MHC molecules and an H-2-linked differentiation antigen as detected by monoclonal antibodies. J Exp Med. 1984 Jan 1;159(1):21–40. doi: 10.1084/jem.159.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Soloski M. J., Hood L., Stroynowski I. Qa-region class I gene expression: identification of a second class I gene, Q9, encoding a Qa-2 polypeptide. Proc Natl Acad Sci U S A. 1988 May;85(9):3100–3104. doi: 10.1073/pnas.85.9.3100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Soloski M. J., Vernachio J., Einhorn G., Lattimore A. Qa gene expression: biosynthesis and secretion of Qa-2 molecules in activated T cells. Proc Natl Acad Sci U S A. 1986 May;83(9):2949–2953. doi: 10.1073/pnas.83.9.2949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Steinmetz M., Moore K. W., Frelinger J. G., Sher B. T., Shen F. W., Boyse E. A., Hood L. A pseudogene homologous to mouse transplantation antigens: transplantation antigens are encoded by eight exons that correlate with protein domains. Cell. 1981 Sep;25(3):683–692. doi: 10.1016/0092-8674(81)90175-6. [DOI] [PubMed] [Google Scholar]
  34. Strominger J. L. The gamma delta T cell receptor and class Ib MHC-related proteins: enigmatic molecules of immune recognition. Cell. 1989 Jun 16;57(6):895–898. doi: 10.1016/0092-8674(89)90326-7. [DOI] [PubMed] [Google Scholar]
  35. Stroynowski I. Molecules related to class-I major histocompatibility complex antigens. Annu Rev Immunol. 1990;8:501–530. doi: 10.1146/annurev.iy.08.040190.002441. [DOI] [PubMed] [Google Scholar]
  36. Stroynowski I., Soloski M., Low M. G., Hood L. A single gene encodes soluble and membrane-bound forms of the major histocompatibility Qa-2 antigen: anchoring of the product by a phospholipid tail. Cell. 1987 Aug 28;50(5):759–768. doi: 10.1016/0092-8674(87)90334-5. [DOI] [PubMed] [Google Scholar]
  37. Ulker N., Hood L. E., Stroynowski I. Molecular signals for phosphatidylinositol modification of the Qa-2 antigen. J Immunol. 1990 Oct 1;145(7):2214–2219. [PubMed] [Google Scholar]
  38. Ulker N., Samuel C. E. Mechanism of interferon action: inhibition of vesicular stomatitis virus replication in human amnion U cells by cloned human gamma-interferon. I. Effect on early and late stages of the viral multiplication cycle. J Biol Chem. 1985 Apr 10;260(7):4319–4323. [PubMed] [Google Scholar]
  39. Vernachio J., Li M., Donnenberg A. D., Soloski M. J. Qa-2 expression in the adult murine thymus. A unique marker for a mature thymic subset. J Immunol. 1989 Jan 1;142(1):48–56. [PubMed] [Google Scholar]
  40. Waneck G. L., Sherman D. H., Calvin S., Allen H., Flavell R. A. Tissue-specific expression of cell-surface Qa-2 antigen from a transfected Q7b gene of C57BL/10 mice. J Exp Med. 1987 May 1;165(5):1358–1370. doi: 10.1084/jem.165.5.1358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Waneck G. L., Sherman D. H., Kincade P. W., Low M. G., Flavell R. A. Molecular mapping of signals in the Qa-2 antigen required for attachment of the phosphatidylinositol membrane anchor. Proc Natl Acad Sci U S A. 1988 Jan;85(2):577–581. doi: 10.1073/pnas.85.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Watts S., Davis A. C., Gaut B., Wheeler C., Hill L., Goodenow R. S. Organization and structure of the Qa genes of the major histocompatibility complex of the C3H mouse: implications for Qa function and class I evolution. EMBO J. 1989 Jun;8(6):1749–1759. doi: 10.1002/j.1460-2075.1989.tb03568.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Zhang H., Scholl R., Browse J., Somerville C. Double stranded DNA sequencing as a choice for DNA sequencing. Nucleic Acids Res. 1988 Feb 11;16(3):1220–1220. doi: 10.1093/nar/16.3.1220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zinkernagel R. M., Doherty P. C. MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness. Adv Immunol. 1979;27:51–177. doi: 10.1016/s0065-2776(08)60262-x. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES