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
. 1995 Apr 11;92(8):3463–3467. doi: 10.1073/pnas.92.8.3463

Molecular cloning and characterization of a cDNA encoding monoclonal nonspecific suppressor factor.

M Nakamura 1, R M Xavier 1, T Tsunematsu 1, Y Tanigawa 1
PMCID: PMC42187  PMID: 7724584

Abstract

The monoclonal nonspecific suppressor factor (MNSF) is a lymphokine product of a murine T-cell hybridoma that inhibits the generation of lipopolysaccharide-induced immunoglobulin-secreting cells in an antigen-nonspecific manner. A cDNA clone encoding MNSF beta (an isoform of MNSF) was isolated and expressed in bacteria. The sequence obtained is virtually identical to the Fau protein, a product of the ubiquitously expressed fau gene with unknown function. Northern blot analysis demonstrated a single, 0.6-kb transcript. Specific polyclonal antibodies against synthetic peptides corresponding to the deduced amino acid sequences were elicited in rabbits. Immunoprecipitation experiments with these antibodies showed that MNSF beta is released extracellularly in an aggregate form, albeit it lacks a signal peptide sequence. The anti-MNSF beta affinity eluate from the MNSF-producing murine hybridoma (E17) and concanavalin A-activated splenocyte culture supernatants inhibited the immunoglobulin production by lipopolysaccharide-activated splenocytes. Recombinant MNSF beta also showed a similar biologic activity. Thus, ubiquitin-like protein(s) may be involved in the regulation of the immune responses.

Full text

PDF
3463

Images in this article

3464Fig. 1
on p.3464
3465Fig. 2
on p.3465
3465Fig. 3
on p.3465
3466Fig. 5
on p.3466

Selected References

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

  1. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gomi H., Tagaya Y., Nakano T., Mikayama T., Ishizaka K. Antigen-binding glycosylation inhibiting factor from a human T-cell hybridoma specific for bee venom phospholipase A2. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2824–2828. doi: 10.1073/pnas.91.7.2824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Green D. R., Bissonnette R., Zheng H. G., Onda T., Echeverri F., Mogil R. J., Steele J. K., Voralia M., Fotedar A. Immunoregulatory activity of the T-cell receptor alpha chain demonstrated by retroviral gene transfer. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8475–8479. doi: 10.1073/pnas.88.19.8475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  5. Gurney M. E., Heinrich S. P., Lee M. R., Yin H. S. Molecular cloning and expression of neuroleukin, a neurotrophic factor for spinal and sensory neurons. Science. 1986 Oct 31;234(4776):566–574. doi: 10.1126/science.3764429. [DOI] [PubMed] [Google Scholar]
  6. Haas A. L., Ahrens P., Bright P. M., Ankel H. Interferon induces a 15-kilodalton protein exhibiting marked homology to ubiquitin. J Biol Chem. 1987 Aug 15;262(23):11315–11323. [PubMed] [Google Scholar]
  7. Hershko A. Ubiquitin-mediated protein degradation. J Biol Chem. 1988 Oct 25;263(30):15237–15240. [PubMed] [Google Scholar]
  8. Iwata M., Katamura K., Kubo R. T., Grey H. M., Ishizaka K. Relationship between T cell receptors and antigen-binding factors. II. Common antigenic determinants and epitope recognition shared by T cell receptors and antigen-binding factors. J Immunol. 1989 Dec 15;143(12):3917–3924. [PubMed] [Google Scholar]
  9. Kas K., Michiels L., Merregaert J. Genomic structure and expression of the human fau gene: encoding the ribosomal protein S30 fused to a ubiquitin-like protein. Biochem Biophys Res Commun. 1992 Sep 16;187(2):927–933. doi: 10.1016/0006-291x(92)91286-y. [DOI] [PubMed] [Google Scholar]
  10. Knight E., Jr, Cordova B. IFN-induced 15-kDa protein is released from human lymphocytes and monocytes. J Immunol. 1991 Apr 1;146(7):2280–2284. [PubMed] [Google Scholar]
  11. Kuchroo V. K., Byrne M. C., Atsumi Y., Greenfield E., Connolly J. B., Whitters M. J., O'Hara R. M., Jr, Collins M., Dorf M. E. T-cell receptor alpha chain plays a critical role in antigen-specific suppressor cell function. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8700–8704. doi: 10.1073/pnas.88.19.8700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Loeb K. R., Haas A. L. The interferon-inducible 15-kDa ubiquitin homolog conjugates to intracellular proteins. J Biol Chem. 1992 Apr 15;267(11):7806–7813. [PubMed] [Google Scholar]
  13. Mikayama T., Nakano T., Gomi H., Nakagawa Y., Liu Y. C., Sato M., Iwamatsu A., Ishii Y., Weiser W. Y., Ishizaka K. Molecular cloning and functional expression of a cDNA encoding glycosylation-inhibiting factor. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10056–10060. doi: 10.1073/pnas.90.21.10056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moore K. W., Vieira P., Fiorentino D. F., Trounstine M. L., Khan T. A., Mosmann T. R. Homology of cytokine synthesis inhibitory factor (IL-10) to the Epstein-Barr virus gene BCRFI. Science. 1990 Jun 8;248(4960):1230–1234. doi: 10.1126/science.2161559. [DOI] [PubMed] [Google Scholar]
  15. Nakamura M., Ogawa H., Tsunematsu T. Characterization of N-terminal amino acid sequence of monoclonal nonspecific suppressor factor. Cell Immunol. 1992 Jan;139(1):139–144. doi: 10.1016/0008-8749(92)90107-z. [DOI] [PubMed] [Google Scholar]
  16. Nakamura M., Ogawa H., Tsunematsu T. Characterization of monoclonal nonspecific suppressor factor (MNSF) with the use of a monoclonal antibody. J Immunol. 1987 Mar 15;138(6):1799–1803. [PubMed] [Google Scholar]
  17. Nakamura M., Ogawa H., Tsunematsu T. IFN-gamma enhances the expression of cell surface receptors for monoclonal nonspecific suppressor factor. Cell Immunol. 1992 Jan;139(1):131–138. doi: 10.1016/0008-8749(92)90106-y. [DOI] [PubMed] [Google Scholar]
  18. Nakamura M., Ogawa H., Tsunematsu T. Isolation and characterization of a monoclonal nonspecific suppressor factor (MNSF) produced by a T cell hybridoma. J Immunol. 1986 Apr 15;136(8):2904–2909. [PubMed] [Google Scholar]
  19. Nakamura M., Xavier R. M., Tanigawa Y. Monoclonal non-specific suppressor factor (MNSF) inhibits the IL4 secretion by bone marrow-derived mast cell (BMMC). FEBS Lett. 1994 Feb 21;339(3):239–242. doi: 10.1016/0014-5793(94)80423-0. [DOI] [PubMed] [Google Scholar]
  20. Olvera J., Wool I. G. The carboxyl extension of a ubiquitin-like protein is rat ribosomal protein S30. J Biol Chem. 1993 Aug 25;268(24):17967–17974. [PubMed] [Google Scholar]
  21. Schlesinger D. H., Goldstein G., Niall H. D. The complete amino acid sequence of ubiquitin, an adenylate cyclase stimulating polypeptide probably universal in living cells. Biochemistry. 1975 May 20;14(10):2214–2218. doi: 10.1021/bi00681a026. [DOI] [PubMed] [Google Scholar]
  22. Tam J. P. Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5409–5413. doi: 10.1073/pnas.85.15.5409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Toniolo D., Persico M., Alcalay M. A "housekeeping" gene on the X chromosome encodes a protein similar to ubiquitin. Proc Natl Acad Sci U S A. 1988 Feb;85(3):851–855. doi: 10.1073/pnas.85.3.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Van Beveren C., Enami S., Curran T., Verma I. M. FBR murine osteosarcoma virus. II. Nucleotide sequence of the provirus reveals that the genome contains sequences acquired from two cellular genes. Virology. 1984 May;135(1):229–243. doi: 10.1016/0042-6822(84)90133-8. [DOI] [PubMed] [Google Scholar]
  25. Xavier R. M., Nakamura M., Tsunematsu T. Isolation and characterization of a human nonspecific suppressor factor from ascitic fluid of systemic lupus erythematosus. Evidence for a human counterpart of the monoclonal nonspecific suppressor factor and relationship to the T cell receptor alpha-chain. J Immunol. 1994 Mar 1;152(5):2624–2632. [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