Skip to main content
PMC home page
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
. 1992 Sep 15;89(18):8681–8685. doi: 10.1073/pnas.89.18.8681

Identification and functional characterization of a TIA-1-related nucleolysin.

A Kawakami 1, Q Tian 1, X Duan 1, M Streuli 1, S F Schlossman 1, P Anderson 1
PMCID: PMC49984  PMID: 1326761

Abstract

We recently reported the molecular cloning of a cytotoxic granule-associated RNA-binding protein designated TIA-1. The ability of recombinant TIA-1 to induce DNA fragmentation in permeabilized cells suggested that this protein is the granule component responsible for inducing apoptosis in cytolytic lymphocyte (CTL) targets. Here we report the characterization of a cDNA encoding a TIA-1-related protein designated TIAR. The deduced amino acid sequence of TIAR reveals it to be a 42-kDa protein possessing three RNA-binding domains and a carboxyl-terminal auxiliary domain. Although the RNA-binding domains of TIA-1 and TIAR share greater than 85% amino acid homology, their carboxyl-terminal auxiliary domains are only 51% homologous. The carboxyl terminus of TIAR contains a lysosome-targeting motif, indicating that TIAR is probably a cytotoxic granule-associated protein. Like TIA-1, purified recombinant TIAR induced DNA fragmentation in permeabilized target cells. Although immunoblotting analysis of post-nuclear supernatants revealed TIA-1 protein to be restricted to CTLs, PCR analysis revealed the expression of TIA-1 and TIAR mRNA transcripts in a wide variety of cell types. Our data suggest that the granules of CTLs contain at least two candidate nucleolysins involved in CTL killing.

Full text

PDF
8681

Images in this article

8682Image
on p.8682
8684Image
on p.8684
8684Image
on p.8684
8684Image
on p.8684

Selected References

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

  1. Allbritton N. L., Verret C. R., Wolley R. C., Eisen H. N. Calcium ion concentrations and DNA fragmentation in target cell destruction by murine cloned cytotoxic T lymphocytes. J Exp Med. 1988 Feb 1;167(2):514–527. doi: 10.1084/jem.167.2.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson P., Nagler-Anderson C., O'Brien C., Levine H., Watkins S., Slayter H. S., Blue M. L., Schlossman S. F. A monoclonal antibody reactive with a 15-kDa cytoplasmic granule-associated protein defines a subpopulation of CD8+ T lymphocytes. J Immunol. 1990 Jan 15;144(2):574–582. [PubMed] [Google Scholar]
  3. Bandziulis R. J., Swanson M. S., Dreyfuss G. RNA-binding proteins as developmental regulators. Genes Dev. 1989 Apr;3(4):431–437. doi: 10.1101/gad.3.4.431. [DOI] [PubMed] [Google Scholar]
  4. Duke R. C., Chervenak R., Cohen J. J. Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6361–6365. doi: 10.1073/pnas.80.20.6361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duke R. C., Persechini P. M., Chang S., Liu C. C., Cohen J. J., Young J. D. Purified perforin induces target cell lysis but not DNA fragmentation. J Exp Med. 1989 Oct 1;170(4):1451–1456. doi: 10.1084/jem.170.4.1451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fukuda M. Lysosomal membrane glycoproteins. Structure, biosynthesis, and intracellular trafficking. J Biol Chem. 1991 Nov 15;266(32):21327–21330. [PubMed] [Google Scholar]
  7. Hameed A., Olsen K. J., Lee M. K., Lichtenheld M. G., Podack E. R. Cytolysis by Ca-permeable transmembrane channels. Pore formation causes extensive DNA degradation and cell lysis. J Exp Med. 1989 Mar 1;169(3):765–777. doi: 10.1084/jem.169.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hayes M. P., Berrebi G. A., Henkart P. A. Induction of target cell DNA release by the cytotoxic T lymphocyte granule protease granzyme A. J Exp Med. 1989 Sep 1;170(3):933–946. doi: 10.1084/jem.170.3.933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henkart P. A. Mechanism of lymphocyte-mediated cytotoxicity. Annu Rev Immunol. 1985;3:31–58. doi: 10.1146/annurev.iy.03.040185.000335. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Martz E., Howell D. M. CTL: virus control cells first and cytolytic cells second? DNA fragmentation, apoptosis and the prelytic halt hypothesis. Immunol Today. 1989 Mar;10(3):79–86. doi: 10.1016/0167-5699(89)90231-4. [DOI] [PubMed] [Google Scholar]
  12. Peters P. J., Borst J., Oorschot V., Fukuda M., Krähenbühl O., Tschopp J., Slot J. W., Geuze H. J. Cytotoxic T lymphocyte granules are secretory lysosomes, containing both perforin and granzymes. J Exp Med. 1991 May 1;173(5):1099–1109. doi: 10.1084/jem.173.5.1099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Podack E. R., Hengartner H., Lichtenheld M. G. A central role of perforin in cytolysis? Annu Rev Immunol. 1991;9:129–157. doi: 10.1146/annurev.iy.09.040191.001021. [DOI] [PubMed] [Google Scholar]
  14. Ritz J., Schmidt R. E., Michon J., Hercend T., Schlossman S. F. Characterization of functional surface structures on human natural killer cells. Adv Immunol. 1988;42:181–211. doi: 10.1016/s0065-2776(08)60845-7. [DOI] [PubMed] [Google Scholar]
  15. Russell J. H. Internal disintegration model of cytotoxic lymphocyte-induced target damage. Immunol Rev. 1983;72:97–118. doi: 10.1111/j.1600-065x.1983.tb01074.x. [DOI] [PubMed] [Google Scholar]
  16. Shi L., Kraut R. P., Aebersold R., Greenberg A. H. A natural killer cell granule protein that induces DNA fragmentation and apoptosis. J Exp Med. 1992 Feb 1;175(2):553–566. doi: 10.1084/jem.175.2.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
  18. Tian Q., Streuli M., Saito H., Schlossman S. F., Anderson P. A polyadenylate binding protein localized to the granules of cytolytic lymphocytes induces DNA fragmentation in target cells. Cell. 1991 Nov 1;67(3):629–639. doi: 10.1016/0092-8674(91)90536-8. [DOI] [PubMed] [Google Scholar]
  19. Trinchieri G. Biology of natural killer cells. Adv Immunol. 1989;47:187–376. doi: 10.1016/S0065-2776(08)60664-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tschopp J., Nabholz M. Perforin-mediated target cell lysis by cytolytic T lymphocytes. Annu Rev Immunol. 1990;8:279–302. doi: 10.1146/annurev.iy.08.040190.001431. [DOI] [PubMed] [Google Scholar]
  21. Tschopp J., Schäfer S., Masson D., Peitsch M. C., Heusser C. Phosphorylcholine acts as a Ca2+-dependent receptor molecule for lymphocyte perforin. Nature. 1989 Jan 19;337(6204):272–274. doi: 10.1038/337272a0. [DOI] [PubMed] [Google Scholar]
  22. Ucker D. S. Cytotoxic T lymphocytes and glucocorticoids activate an endogenous suicide process in target cells. Nature. 1987 May 7;327(6117):62–64. doi: 10.1038/327062a0. [DOI] [PubMed] [Google Scholar]
  23. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]
  24. Young J. D., Liu C. C. Multiple mechanisms of lymphocyte-mediated killing. Immunol Today. 1988 May;9(5):140–144. doi: 10.1016/0167-5699(88)91201-7. [DOI] [PubMed] [Google Scholar]
  25. Young J. D., Nathan C. F., Podack E. R., Palladino M. A., Cohn Z. A. Functional channel formation associated with cytotoxic T-cell granules. Proc Natl Acad Sci U S A. 1986 Jan;83(1):150–154. doi: 10.1073/pnas.83.1.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yue C. C., Reynolds C. W., Henkart P. A. Inhibition of cytolysin activity in large granular lymphocyte granules by lipids: evidence for a membrane insertion mechanism of lysis. Mol Immunol. 1987 Jun;24(6):647–653. doi: 10.1016/0161-5890(87)90046-0. [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