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J Exp Med. 1994 March 1; 179(3): 901–909.
PMCID: PMC2191397

Interferon gamma stimulation modulates the proteolytic activity and cleavage site preference of 20S mouse proteasomes


The proteasome is a 700-kD multisubunit enzyme complex with several proteolytically active sites. The enzyme complex is involved in both ubiquitin-dependent and -independent protein degradation and may contribute to the processing of antigens presented by major histocompatibility complex (MHC) class I molecules. Here we demonstrate that treatment of mouse fibroblast cells with 20 U interferon gamma (IFN-gamma) for 3 d induces a change in the proteasome subunit composition and that the beta-type subunit LMP2, which is encoded in the MHC class II region, is incorporated into the enzyme complex. This is paralleled by reduction of the homologous delta-subunit. IFN-gamma stimulation results in a downregulation of the chymotrypsin-like Suc- LLVY-MCA peptide hydrolyzing activity of 20S proteasomes whereas the trypsin-like activity remains unaffected. When tested as a substrate a synthetic 25-mer polypeptide whose sequence covers the antigenic nonapeptide YPHFMPTNL of the MCMV pp89, 20S proteasomes of IFN-gamma- induced cells exhibit altered chymotrypsin-like cleavage site preferences. In the absence of IFN-gamma induction, the naturally processed nonamer peptide that is presented by MHC class I molecules appears as a minor cleavage product. IFN-gamma activation does not result in an increase of the final peptide but results in a different set of peptides. We hypothesize that these peptides represent precursor peptides that can be trimmed to final peptide size.

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Selected References

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  • Goldberg AL. The mechanism and functions of ATP-dependent proteases in bacterial and animal cells. Eur J Biochem. 1992 Jan 15;203(1-2):9–23. [PubMed]
  • Rivett AJ. Proteasomes: multicatalytic proteinase complexes. Biochem J. 1993 Apr 1;291(Pt 1):1–10. [PubMed]
  • Dahlmann B, Rutschmann M, Kuehn L, Reinauer H. Activation of the multicatalytic proteinase from rat skeletal muscle by fatty acids or sodium dodecyl sulphate. Biochem J. 1985 May 15;228(1):171–177. [PubMed]
  • Orlowski M. The multicatalytic proteinase complex, a major extralysosomal proteolytic system. Biochemistry. 1990 Nov 13;29(45):10289–10297. [PubMed]
  • Orlowski M. The multicatalytic proteinase complex (proteasome) and intracellular protein degradation: diverse functions of an intracellular particle. J Lab Clin Med. 1993 Feb;121(2):187–189. [PubMed]
  • McGuire MJ, McCullough ML, Croall DE, DeMartino GN. The high molecular weight multicatalytic proteinase, macropain, exists in a latent form in human erythrocytes. Biochim Biophys Acta. 1989 Apr 6;995(2):181–186. [PubMed]
  • Kleinschmidt JA, Hügle B, Grund C, Franke WW. The 22 S cylinder particles of Xenopus laevis. I. Biochemical and electron microscopic characterization. Eur J Cell Biol. 1983 Nov;32(1):143–156. [PubMed]
  • Baumeister W, Dahlmann B, Hegerl R, Kopp F, Kuehn L, Pfeifer G. Electron microscopy and image analysis of the multicatalytic proteinase. FEBS Lett. 1988 Dec 5;241(1-2):239–245. [PubMed]
  • Tanaka K, Tamura T, Yoshimura T, Ichihara A. Proteasomes: protein and gene structures. New Biol. 1992 Mar;4(3):173–187. [PubMed]
  • Kloetzel PM, Frentzel S, Gernold M, Haass C, Klein U, Pesold-Hurt B, Seelig A. The proteasome of Drosophila and features of the evolutionarily conserved PROS-gene family. Biomed Biochim Acta. 1991;50(4-6):451–457. [PubMed]
  • Heinemeyer W, Gruhler A, Möhrle V, Mahé Y, Wolf DH. PRE2, highly homologous to the human major histocompatibility complex-linked RING10 gene, codes for a yeast proteasome subunit necessary for chrymotryptic activity and degradation of ubiquitinated proteins. J Biol Chem. 1993 Mar 5;268(7):5115–5120. [PubMed]
  • Zwickl P, Lottspeich F, Baumeister W. Expression of functional Thermoplasma acidophilum proteasomes in Escherichia coli. FEBS Lett. 1992 Nov 9;312(2-3):157–160. [PubMed]
  • Seufert W, Jentsch S. In vivo function of the proteasome in the ubiquitin pathway. EMBO J. 1992 Aug;11(8):3077–3080. [PubMed]
  • Richter-Ruoff B, Heinemeyer W, Wolf DH. The proteasome/multicatalytic-multifunctional proteinase. In vivo function in the ubiquitin-dependent N-end rule pathway of protein degradation in eukaryotes. FEBS Lett. 1992 May 11;302(2):192–196. [PubMed]
  • Shimbara N, Orino E, Sone S, Ogura T, Takashina M, Shono M, Tamura T, Yasuda H, Tanaka K, Ichihara A. Regulation of gene expression of proteasomes (multi-protease complexes) during growth and differentiation of human hematopoietic cells. J Biol Chem. 1992 Sep 5;267(25):18100–18109. [PubMed]
  • Klein U, Gernold M, Kloetzel PM. Cell-specific accumulation of Drosophila proteasomes (MCP) during early development. J Cell Biol. 1990 Dec;111(6 Pt 1):2275–2282. [PMC free article] [PubMed]
  • Glynne R, Powis SH, Beck S, Kelly A, Kerr LA, Trowsdale J. A proteasome-related gene between the two ABC transporter loci in the class II region of the human MHC. Nature. 1991 Sep 26;353(6342):357–360. [PubMed]
  • Kelly A, Powis SH, Glynne R, Radley E, Beck S, Trowsdale J. Second proteasome-related gene in the human MHC class II region. Nature. 1991 Oct 17;353(6345):667–668. [PubMed]
  • Ortiz-Navarrete V, Seelig A, Gernold M, Frentzel S, Kloetzel PM, Hämmerling GJ. Subunit of the '20S' proteasome (multicatalytic proteinase) encoded by the major histocompatibility complex. Nature. 1991 Oct 17;353(6345):662–664. [PubMed]
  • Brown MG, Driscoll J, Monaco JJ. Structural and serological similarity of MHC-linked LMP and proteasome (multicatalytic proteinase) complexes. Nature. 1991 Sep 26;353(6342):355–357. [PubMed]
  • Martinez CK, Monaco JJ. Homology of proteasome subunits to a major histocompatibility complex-linked LMP gene. Nature. 1991 Oct 17;353(6345):664–667. [PubMed]
  • Yang Y, Waters JB, Früh K, Peterson PA. Proteasomes are regulated by interferon gamma: implications for antigen processing. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4928–4932. [PubMed]
  • Früh K, Yang Y, Arnold D, Chambers J, Wu L, Waters JB, Spies T, Peterson PA. Alternative exon usage and processing of the major histocompatibility complex-encoded proteasome subunits. J Biol Chem. 1992 Nov 5;267(31):22131–22140. [PubMed]
  • Rötzschke O, Falk K. Naturally-occurring peptide antigens derived from the MHC class-I-restricted processing pathway. Immunol Today. 1991 Dec;12(12):447–455. [PubMed]
  • Michalek MT, Grant EP, Gramm C, Goldberg AL, Rock KL. A role for the ubiquitin-dependent proteolytic pathway in MHC class I-restricted antigen presentation. Nature. 1993 Jun 10;363(6429):552–554. [PubMed]
  • Arnold D, Driscoll J, Androlewicz M, Hughes E, Cresswell P, Spies T. Proteasome subunits encoded in the MHC are not generally required for the processing of peptides bound by MHC class I molecules. Nature. 1992 Nov 12;360(6400):171–174. [PubMed]
  • Momburg F, Ortiz-Navarrete V, Neefjes J, Goulmy E, van de Wal Y, Spits H, Powis SJ, Butcher GW, Howard JC, Walden P, et al. Proteasome subunits encoded by the major histocompatibility complex are not essential for antigen presentation. Nature. 1992 Nov 12;360(6400):174–177. [PubMed]
  • Kloetzel PM, Falkenburg PE, Hössl P, Glätzer KH. The 19S ring-type particles of Drosophila. Cytological and biochemical analysis of their intracellular association and distribution. Exp Cell Res. 1987 May;170(1):204–213. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • O'Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed]
  • Seelig A, Kloetzel PM, Kuehn L, Dahlmann B. Molecular interaction of the proteasome (multicatalytic proteinase). Evidence that the proteasome is not a constituent of the '26 S' multienzyme complex. Biochem J. 1991 Nov 15;280(Pt 1):225–232. [PubMed]
  • Reddehase MJ, Rothbard JB, Koszinowski UH. A pentapeptide as minimal antigenic determinant for MHC class I-restricted T lymphocytes. Nature. 1989 Feb 16;337(6208):651–653. [PubMed]
  • Del Val M, Schlicht HJ, Ruppert T, Reddehase MJ, Koszinowski UH. Efficient processing of an antigenic sequence for presentation by MHC class I molecules depends on its neighboring residues in the protein. Cell. 1991 Sep 20;66(6):1145–1153. [PubMed]
  • DeMartino GN, Orth K, McCullough ML, Lee LW, Munn TZ, Moomaw CR, Dawson PA, Slaughter CA. The primary structures of four subunits of the human, high-molecular-weight proteinase, macropain (proteasome), are distinct but homologous. Biochim Biophys Acta. 1991 Aug 9;1079(1):29–38. [PubMed]
  • Frentzel S, Kuhn-Hartmann I, Gernold M, Gött P, Seelig A, Kloetzel PM. The major-histocompatibility-complex-encoded beta-type proteasome subunits LMP2 and LMP7. Evidence that LMP2 and LMP7 are synthesized as proproteins and that cellular levels of both mRNA and LMP-containing 20S proteasomes are differentially regulated. Eur J Biochem. 1993 Aug 15;216(1):119–126. [PubMed]
  • Zwickl P, Grziwa A, Pühler G, Dahlmann B, Lottspeich F, Baumeister W. Primary structure of the Thermoplasma proteasome and its implications for the structure, function, and evolution of the multicatalytic proteinase. Biochemistry. 1992 Feb 4;31(4):964–972. [PubMed]
  • Haass C, Kloetzel PM. The Drosophila proteasome undergoes changes in its subunit pattern during development. Exp Cell Res. 1989 Jan;180(1):243–252. [PubMed]
  • Reddehase MJ, Koszinowski UH. Redistribution of critical major histocompatibility complex and T cell receptor-binding functions of residues in an antigenic sequence after biterminal substitution. Eur J Immunol. 1991 Jul;21(7):1697–1701. [PubMed]
  • Driscoll J, Brown MG, Finley D, Monaco JJ. MHC-linked LMP gene products specifically alter peptidase activities of the proteasome. Nature. 1993 Sep 16;365(6443):262–264. [PubMed]
  • Gaczynska M, Rock KL, Goldberg AL. Gamma-interferon and expression of MHC genes regulate peptide hydrolysis by proteasomes. Nature. 1993 Sep 16;365(6443):264–267. [PubMed]

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