γ-Interferon transcriptionally regulates an early-response gene containing homology to platelet proteins

Interferons are a family of proteins first identified by their ability to induce cellular resistance to infection by many viruses. In addition to the antiviral properties it shares with the α- and β-interferons, γ-interferon (IFN-γ), a lymphokine secreted by activated T cells, activates macrophages, stimulates B cells, increases fibroblast and endothelial cell resistance to many non-viral intracellular parasites and modulates cell-surface proteins central to immune cell regulation1–13. To identify molecules involved in the IFN-γ response and characterize their modulation, we have isolated genes that are induced following recombinant IFN-γ treatment of U937 cells, a histiocytic lymphoma cell line with monocytic characteristics14,15. We report here the molecular cloning and characterization of a gene regulated by rIFN-γ in U937 cells as well as in human mononuclear cells, fibroblasts and endothelial cells. Messenger RNA from this gene is induced within 30 min of rIFN-γ treatment and demonstrates maximal (>30-fold) accumulation within 5 h. Increased transcription is partly responsible for this accumulation. This gene encodes a protein of relative molecular mass (Mr) 12,378 which has significant amino-acid homology to platelet factor-4 and β-thromboglobulin, two chemo-tatic proteins released by platelets on degranulation. This IFN-γ-inducible protein may be a member of a family of proteins involved in the inflammatory process.

[1]  D. Kemp,et al.  Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[2]  H. Winkler,et al.  Cloned mouse interferon-gamma inhibits the growth of Rickettsia prowazekii in cultured mouse fibroblasts , 1983, The Journal of experimental medicine.

[3]  S. Lonky,et al.  Stimulation of human leukocyte elastase by platelet factor 4. Physiologic, morphologic, and biochemical effects on hamster lungs in vitro. , 1981, The Journal of clinical investigation.

[4]  D Perlman,et al.  A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. , 1983, Journal of molecular biology.

[5]  M. Drillings,et al.  Platelet alpha-granule proteins: studies on release and subcellular localization , 1979 .

[6]  G. Ringold.  Glucocorticoid regulation of mouse mammary tumor virus gene expression. , 1979, Biochimica et biophysica acta.

[7]  K. Nilsson,et al.  Establishment and characterization of a human histiocytic lymphoma cell line (U‐937) , 1976, International journal of cancer.

[8]  D. Lipman,et al.  Rapid and sensitive protein similarity searches. , 1985, Science.

[9]  F. Sanger,et al.  A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. , 1975, Journal of molecular biology.

[10]  H. Okayama,et al.  High-efficiency cloning of full-length cDNA , 1982, Molecular and cellular biology.

[11]  W. Fiers,et al.  Immune interferon activates multiple class II major histocompatibility complex genes and the associated invariant chain gene in human endothelial cells and dermal fibroblasts. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[12]  E. Goetzl,et al.  Stimulation of histamine release from human basophils by human platelet factor 4. , 1983, The Journal of clinical investigation.

[13]  J. Vilček,et al.  Activation of human monocyte cytotoxicity by natural and recombinant immune interferon. , 1983, Journal of immunology.

[14]  W. Gilbert,et al.  A new method for sequencing DNA. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[15]  T. Martin,et al.  Human β-thromboglobulin inhibits PGI2 production and binds to a specific site in bovine aortic endothelial cells , 1979, Nature.

[16]  H. Cohen,et al.  Purification and binding properties of human platelet factor four. , 1976, The Journal of biological chemistry.

[17]  S. B. Needleman,et al.  A general method applicable to the search for similarities in the amino acid sequence of two proteins. , 1970, Journal of molecular biology.

[18]  J. D. Marshall,et al.  γ-Interferon is one of several direct B cell-maturing lymphokines , 1984, Nature.

[19]  P. Marrack,et al.  Role of γ-interferon in antibody-producing responses , 1984, Nature.

[20]  M. Fellous,et al.  Preferential effect of γ interferon on the synthesis of HLA antigens and their mRNAs in human cells , 1982, Nature.

[21]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[22]  S. Niewiarowski,et al.  Conversion of low-affinity platelet factor 4 to beta-thromboglobulin by plasmin and trypsin. , 1980, Biochimica et biophysica acta.

[23]  J. Darnell,et al.  Transcriptional induction of two genes in human cells by beta interferon. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[24]  N. Hay,et al.  Control of late simian virus 40 transcription by the attenuation mechanism and transcriptionally active ternary complexes are associated with the nuclear matrix. , 1984, Journal of molecular biology.

[25]  J. Larrick,et al.  In vitro activation of a human macrophage-like cell line , 1979, Nature.

[26]  H. Murray,et al.  Killing of intracellular Leishmania donovani by lymphokine-stimulated human mononuclear phagocytes. Evidence that interferon-gamma is the activating lymphokine. , 1983, The Journal of clinical investigation.

[27]  J. Seidman,et al.  Mouse beta 2-microglobulin cDNA clones: a screening procedure for cDNA clones corresponding to rare mRNAs. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Gunnar von Heijne,et al.  Patterns of Amino Acids near Signal‐Sequence Cleavage Sites , 1983 .

[29]  S. Russell,et al.  Recombinant mouse gamma interferon induces the priming step in macrophage activation for tumor cell killing. , 1983, Journal of immunology.

[30]  R. Doolittle,et al.  A simple method for displaying the hydropathic character of a protein. , 1982, Journal of molecular biology.

[31]  G. Stark,et al.  Transcriptional and posttranscriptional regulation of interferon-induced gene expression in human cells , 1984, Cell.

[32]  T. Merigan,et al.  Recombinant interferon-gamma increases HLA-DR synthesis and expression. , 1983, Journal of immunology.

[33]  D. Pepper,et al.  Complete covalent structure of human beta-thromboglobulin. , 1978, Biochemistry.

[34]  J. W. Miller,et al.  Structural and biological characteristics of connective tissue activating peptide (CTAP-III), a major human platelet-derived growth factor. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[35]  R. Schultz,et al.  Functional identity between murine γ Interferon and macrophage activating factor , 1983, Nature.

[36]  E. Pfefferkorn.  Interferon gamma blocks the growth of Toxoplasma gondii in human fibroblasts by inducing the host cells to degrade tryptophan. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[37]  P. Lomedico,et al.  A bacterial clone synthesizing proinsulin. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[38]  W. Bodmer,et al.  Sequence of an HLA-DR α-chain cDNA clone and intron-exon organization of the corresponding gene , 1982, Nature.

[39]  S. Weissman,et al.  Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[40]  R. Schreiber,et al.  Evidence for a gamma-interferon receptor that regulates macrophage tumoricidal activity , 1984, The Journal of experimental medicine.

[41]  J. Darnell,et al.  The primary transcription unit of the mouse β-Major globin gene , 1981, Cell.

[42]  F C Kafatos,et al.  Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. , 1979, Nucleic acids research.

[43]  R. Senior,et al.  Chemotactic activity of platelet alpha granule proteins for fibroblasts , 1983, The Journal of cell biology.

[44]  R. Heinrikson,et al.  Platelet factor 4 is chemotactic for neutrophils and monocytes. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[45]  C. Nathan,et al.  Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity , 1983, The Journal of experimental medicine.

[46]  E. Harper,et al.  Platelet factor 4: an inhibitor of collagenase. , 1978, Science.

[47]  T. Manser,et al.  Effect of IFN-γ on the immune response in vivo and on gene expression in vitro , 1984, Nature.