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Cycloheximide, an inhibitor of protein synthesis, prevents γ-interferon-induced expression of class II mRNA in a macrophage cell line

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Abstract

To characterize the mechanisms by which interferon γ (IFN-γ) upregulates major histocompatibility complex class II mRNA levels in mouse macrophages, we studied the effect of IFN-γ on the transcription rate of class II genes and investigated the requirement for ongoing protein synthesis for the induction of class II mRNA expression. Nuclear run-off assays demonstrate that IFN-γ induces class II mRNA at the transcriptional level. Treatment with cycloheximide, an inhibitor of protein synthesis, prevented the IFN-γ-mediated accumulation of E α mRNA in the mouse macrophage cell line P388 D.1, indicating that induction of E α mRNA in P388 D.1 cells requires de novo synthesis of a protein intermediate. Our studies suggest that this putative protein factor is labile and required throughout the induction period.

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Abbreviations

IFN-γ:

interferon=γ

CHX:

cycloheximide

MHC:

major histocompatibility complex

PK:

pyruvate kinase

References

  • Accolla, R. S., Scarpellino, L., Carra, G., and Guardiola, J.: Transacting element(s) operating across barriers positively regulate expression of the major histocompatibility complex class II genes. J. Exp. Med. 162: 1117–1133, 1985

    Google Scholar 

  • Benoist, C. O., Mathis, D. J., Kanter, M. R., Williams, V. E., and McDevitt, H. O.: The murine la a chains, Eα and Aα, show a surprising degree of sequence homology. Proc. Natl. Acad. Sci. U.S.A. 80: 534–538, 1983

    Google Scholar 

  • Dani, C. H., Blanchard, J. M., Piechaczyk, M., Saborty, S. E., Marty, L., and Jeanteur, P.: Extreme instability of myc RNA in normal and transformed human cells. Proc. Natl. Acad. Sci. U.S.A. 81: 7046–7050, 1984

    Google Scholar 

  • Fertsch, D., Shoenberg, D. R., Germain, R. N., Ton, J. Y. L., and Vogel, S. N.: Induction of macrophage la antigen expression by rIFNγ and down-regulation by IFNα/β and dexamethasone are mediated by changes in steady-state levels of Ia mRNA. J. Immunol. 139: 244–249, 1987

    Google Scholar 

  • Flavell, R. A., Allen, H., Burkly, L. C., Sherman, D. H., Waneck, G. L., and Widera, G.: Molecular biology of the H-2 histocompatibility complex. Science 233: 437–443, 1986

    Google Scholar 

  • Friedman, R. L., Manly, S. P., McMahon, M., Kerr, I. M., and Stark, G. R.: Transcriptional and post-transcriptional regulation of interferon-induced gene expression in human cells. Cell 38: 745–755, 1984

    Google Scholar 

  • Greenberg, H. E. and Ziff, E. B.: Stimulation of 3173 cells induces transcription of the c-fos proto-oncogene. Nature 311: 433–438, 1984

    Google Scholar 

  • Hyldig-Nielsen, J. J., Schenning, L., Hämmerling, D., Widmark, E., Helden, E., Lind, P., Servenius, B., Lund, T., Flavell, R., Lee, J. S., Trowsdale, J., Schreier, P. M., Zablitsky, F., Larhammar, D., Peterson, P. A., and Rask, L.: The complete nucleotide sequence of I-E α d immune response gene. Nucleic Acids Res. 11: 5055–5071, 1983

    Google Scholar 

  • King, D. P., Sizer, K. C., Freund, Y. R., Jones, P. P., and Parnes, J. R.: Coordinate induction of Ia α, β, and 1i mRNA in a macrophage cell line. J. Immunol. 135: 632–636, 1985

    Google Scholar 

  • Koren, H. S., Handwerger, B. S., and Wunderlich, J. R.: Identification of macrophage-like characteristics in a cultured murine tumor line. J. Immunol. 114: 894–897, 1975

    Google Scholar 

  • LaLanne, J.-L., Delarbre, C., Gachelin, G., and Kourilsky, P.: A cDNA clone containing the entire coding sequence of a mouse H-2Kd histocompatibility antigen. Nucleic Acids Res. 11: 1567–1577, 1983

    Google Scholar 

  • Larhammar, D., Hämmerling, U., Denaro, M., Lund, T., Flavell, R. A., Rask, L., and Peterson, P. A.: Structure of the murine I-A β locus: sequence of the I-A Inβ gene and an adjacent β-chain second domain exon. Cell 34: 179–188, 1983

    Google Scholar 

  • Lee, K.-C.: Regulation of T-cell activation by macrophage subsets. In S. B. Mizel (ed.) Lymphokines, Volume 6, pp. 1–23, Academic Press, New York, 1982

    Google Scholar 

  • Lewis, S. A., Gilmartin, M. E., Hall, J. L., and Cowan, N. J.: Three expressed sequences within the human β-tubulin multigene family each define a distinct isotype. J. Mol. Biol. 182: 11–20, 1985

    Google Scholar 

  • Linial, M., Gunderson, N., and Groudine, M.: Enhanced transcription of c-myc in bsaal lymphoma cells requires continuous protein synthesis. Science 230: 1126–1132, 1985

    Google Scholar 

  • Maniatis, T., Fritsch, E., and Sambrook, J.: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1982

    Google Scholar 

  • Revel, M. and Chebath, J.: Interferon-activated genes. Trends Biochem. Sci. 11: 166–170, 1986

    Google Scholar 

  • Ringold, G. M.: Glucocorticoid regulation of mouse mammary tumor virus gene expression. Biochim. Biophys. Acta 560: 487–508, 1979

    Google Scholar 

  • Ringold, G. M., Dieckmann, B., Vannice, J. L., Trahey, M., and McCormick, F.: Inhibition of protein synthesis stimulates the transcription of human β-interferon genes in Chinese hamster ovary cells. Proc. Natl. Acad. Sci. U.S.A. 81: 3964–3968, 1984

    Google Scholar 

  • Sen, R. and Baltimore, D.: Inducibility of κ immunoglobulin enhancer-binding protein NF-κB by a post-translational mechanism. Cell 47: 921–928, 1986

    Google Scholar 

  • Steinman, R. M., Nogueira, N., Witmer, M. D., Tydings, J. G., and Mellmann, I. S.: Lymphokine enhances the expression and synthesis of la antigens on cultured mouse peritoneal macrophages J. Exp. Med. 152: 1248–1261, 1980

    Google Scholar 

  • Unanue, E. R. and Allen, P. M.: The basis for the immunoregulatory role of macrophages and other accessory cells. Science 236: 551–557, 1987

    Google Scholar 

  • Varesio, L.: Molecular bases for macrophage activation. Ann. Inst. Pasteur 137C: 235–240, 1986

    Google Scholar 

  • Walker, E. B., Maino, V., Sanchez-Lanier, M., Warner, N., Stewart, C.: Murine gamma interferon activates the release of a macrophage-derived Ia-inducing factor that transfers la inductive capacity. J. Exp. Med. 159: 1532–1547, 1984

    Google Scholar 

  • Wall, R., Briskin, M., Carter, C., Govan, H., Taylor, A., and Kincade, P.: A labile inhibitor blocks immunoglobulin κ-light-chain-gene transcription in a pre-B leukemic cell line. Proc. Natl. Acad. Sci. U.S.A. 83: 295–298, 1986

    Google Scholar 

  • Widera, G. and Flavell, R. A.: The nucleotide sequence of the murine I-E β d immune response gene: evidence for gene conversion events in class II genes of the major histocompatibility complex. EMBO J. 3: 1221–1125, 1984

    Google Scholar 

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Author notes

  1. Erik C. Böttger & Richard A. Flavell

    Present address: Institut für medizinische Mikrobiologie, Medizinische Hochschule Hannover, Konstantyn-Gutscho-Str. 8, 3000, Hannover 61, FRG

  2. Michael A. Blanar

    Present address: Section for Immunobiology and Howard Hughes Medical Institute, Yale University School of Medicine, 310 Cedar St., 06510, New Medical Institute, CT, New Haven, USA

Authors and Affiliations

  1. Biogen Research Corp., 14 Cambridge Center, 02142, MA, Cambridge, USA

    Erik C. Böttger, Michael A. Blanar & Richard A. Flavell

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  1. Erik C. Böttger
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  2. Michael A. Blanar
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  3. Richard A. Flavell
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denotes that this paper resulted from an equal contribution by the first two authors

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Böttger, E.C., Blanar, M.A. & Flavell, R.A. Cycloheximide, an inhibitor of protein synthesis, prevents γ-interferon-induced expression of class II mRNA in a macrophage cell line. Immunogenetics 28, 215–220 (1988). https://doi.org/10.1007/BF00345497

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  • DOI: https://doi.org/10.1007/BF00345497

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