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. 1993 Jan;13(1):276–288. doi: 10.1128/mcb.13.1.276

Acute-phase response factor, a nuclear factor binding to acute-phase response elements, is rapidly activated by interleukin-6 at the posttranslational level.

U M Wegenka 1, J Buschmann 1, C Lütticken 1, P C Heinrich 1, F Horn 1
PMCID: PMC358907  PMID: 7678052

Abstract

Interleukin-6 (IL-6) is known to be a major mediator of the acute-phase response in liver. We show here that IL-6 triggers the rapid activation of a nuclear factor, termed acute-phase response factor (APRF), both in rat liver in vivo and in human hepatoma (HepG2) cells in vitro. APRF bound to IL-6 response elements in the 5'-flanking regions of various acute-phase protein genes (e.g., the alpha 2-macroglobulin, fibrinogen, and alpha 1-acid glycoprotein genes). These elements contain a characteristic hexanucleotide motif, CTGGGA, known to be required for the IL-6 responsiveness of these genes. Analysis of the binding specificity of APRF revealed that it is different from NF-IL6 and NF-kappa B, transcription factors known to be regulated by cytokines and involved in the transcriptional regulation of acute-phase protein genes. In HepG2 cells, activation of APRF was observed within minutes after stimulation with IL-6 or leukemia-inhibitory factor and did not require ongoing protein synthesis. Therefore, a preexisting inactive form of APRF is activated by a posttranslational mechanism. We present evidence that this activation occurs in the cytoplasm and that a phosphorylation is involved. These results lead to the conclusions that APRF is an immediate target of the IL-6 signalling cascade and is likely to play a central role in the transcriptional regulation of many IL-6-induced genes.

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Selected References

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

  1. Akira S., Isshiki H., Sugita T., Tanabe O., Kinoshita S., Nishio Y., Nakajima T., Hirano T., Kishimoto T. A nuclear factor for IL-6 expression (NF-IL6) is a member of a C/EBP family. EMBO J. 1990 Jun;9(6):1897–1906. doi: 10.1002/j.1460-2075.1990.tb08316.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amin J., Ananthan J., Voellmy R. Key features of heat shock regulatory elements. Mol Cell Biol. 1988 Sep;8(9):3761–3769. doi: 10.1128/mcb.8.9.3761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andus T., Geiger T., Hirano T., Kishimoto T., Tran-Thi T. A., Decker K., Heinrich P. C. Regulation of synthesis and secretion of major rat acute-phase proteins by recombinant human interleukin-6 (BSF-2/IL-6) in hepatocyte primary cultures. Eur J Biochem. 1988 Apr 15;173(2):287–293. doi: 10.1111/j.1432-1033.1988.tb13997.x. [DOI] [PubMed] [Google Scholar]
  4. Baldwin A. S., Jr, Sharp P. A. Binding of a nuclear factor to a regulatory sequence in the promoter of the mouse H-2Kb class I major histocompatibility gene. Mol Cell Biol. 1987 Jan;7(1):305–313. doi: 10.1128/mcb.7.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bao J. J., Sifers R. N., Kidd V. J., Ledley F. D., Woo S. L. Molecular evolution of serpins: homologous structure of the human alpha 1-antichymotrypsin and alpha 1-antitrypsin genes. Biochemistry. 1987 Dec 1;26(24):7755–7759. doi: 10.1021/bi00398a033. [DOI] [PubMed] [Google Scholar]
  6. Baumann H., Jahreis G. P., Morella K. K. Interaction of cytokine- and glucocorticoid-response elements of acute-phase plasma protein genes. Importance of glucocorticoid receptor level and cell type for regulation of the elements from rat alpha 1-acid glycoprotein and beta-fibrinogen genes. J Biol Chem. 1990 Dec 25;265(36):22275–22281. [PubMed] [Google Scholar]
  7. Baumann H., Jahreis G. P., Morella K. K., Won K. A., Pruitt S. C., Jones V. E., Prowse K. R. Transcriptional regulation through cytokine and glucocorticoid response elements of rat acute phase plasma protein genes by C/EBP and JunB. J Biol Chem. 1991 Oct 25;266(30):20390–20399. [PubMed] [Google Scholar]
  8. Baumann H., Prowse K. R., Marinković S., Won K. A., Jahreis G. P. Stimulation of hepatic acute phase response by cytokines and glucocorticoids. Ann N Y Acad Sci. 1989;557:280-95, discussion 295-6. doi: 10.1111/j.1749-6632.1989.tb24021.x. [DOI] [PubMed] [Google Scholar]
  9. Baumann H., Won K. A., Jahreis G. P. Human hepatocyte-stimulating factor-III and interleukin-6 are structurally and immunologically distinct but regulate the production of the same acute phase plasma proteins. J Biol Chem. 1989 May 15;264(14):8046–8051. [PubMed] [Google Scholar]
  10. Baumann H., Wong G. G. Hepatocyte-stimulating factor III shares structural and functional identity with leukemia-inhibitory factor. J Immunol. 1989 Aug 15;143(4):1163–1167. [PubMed] [Google Scholar]
  11. Beato M., Chalepakis G., Schauer M., Slater E. P. DNA regulatory elements for steroid hormones. J Steroid Biochem. 1989 May;32(5):737–747. doi: 10.1016/0022-4731(89)90521-9. [DOI] [PubMed] [Google Scholar]
  12. Bohmann D. Transcription factor phosphorylation: a link between signal transduction and the regulation of gene expression. Cancer Cells. 1990 Nov;2(11):337–344. [PubMed] [Google Scholar]
  13. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  14. Busch S. J., Sassone-Corsi P. Dimers, leucine zippers and DNA-binding domains. Trends Genet. 1990 Feb;6(2):36–40. doi: 10.1016/0168-9525(90)90071-d. [DOI] [PubMed] [Google Scholar]
  15. Chang C. J., Chen T. T., Lei H. Y., Chen D. S., Lee S. C. Molecular cloning of a transcription factor, AGP/EBP, that belongs to members of the C/EBP family. Mol Cell Biol. 1990 Dec;10(12):6642–6653. doi: 10.1128/mcb.10.12.6642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Descombes P., Chojkier M., Lichtsteiner S., Falvey E., Schibler U. LAP, a novel member of the C/EBP gene family, encodes a liver-enriched transcriptional activator protein. Genes Dev. 1990 Sep;4(9):1541–1551. doi: 10.1101/gad.4.9.1541. [DOI] [PubMed] [Google Scholar]
  17. Edbrooke M. R., Foldi J., Cheshire J. K., Li F., Faulkes D. J., Woo P. Constitutive and NF-kappa B-like proteins in the regulation of the serum amyloid A gene by interleukin 1. Cytokine. 1991 Sep;3(5):380–388. doi: 10.1016/1043-4666(91)90041-b. [DOI] [PubMed] [Google Scholar]
  18. Flohé S., Heinrich P. C., Schneider J., Wendel A., Flohé L. Time course of IL-6 and TNF alpha release during endotoxin-induced endotoxin tolerance in rats. Biochem Pharmacol. 1991 Jun 1;41(11):1607–1614. doi: 10.1016/0006-2952(91)90161-w. [DOI] [PubMed] [Google Scholar]
  19. Fowlkes D. M., Mullis N. T., Comeau C. M., Crabtree G. R. Potential basis for regulation of the coordinately expressed fibrinogen genes: homology in the 5' flanking regions. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2313–2316. doi: 10.1073/pnas.81.8.2313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ganter U., Arcone R., Toniatti C., Morrone G., Ciliberto G. Dual control of C-reactive protein gene expression by interleukin-1 and interleukin-6. EMBO J. 1989 Dec 1;8(12):3773–3779. doi: 10.1002/j.1460-2075.1989.tb08554.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Gauldie J., Richards C., Harnish D., Lansdorp P., Baumann H. Interferon beta 2/B-cell stimulatory factor type 2 shares identity with monocyte-derived hepatocyte-stimulating factor and regulates the major acute phase protein response in liver cells. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7251–7255. doi: 10.1073/pnas.84.20.7251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gearing D. P., Comeau M. R., Friend D. J., Gimpel S. D., Thut C. J., McGourty J., Brasher K. K., King J. A., Gillis S., Mosley B. The IL-6 signal transducer, gp130: an oncostatin M receptor and affinity converter for the LIF receptor. Science. 1992 Mar 13;255(5050):1434–1437. doi: 10.1126/science.1542794. [DOI] [PubMed] [Google Scholar]
  24. Geiger T., Andus T., Klapproth J., Hirano T., Kishimoto T., Heinrich P. C. Induction of rat acute-phase proteins by interleukin 6 in vivo. Eur J Immunol. 1988 May;18(5):717–721. doi: 10.1002/eji.1830180510. [DOI] [PubMed] [Google Scholar]
  25. Ghosh S., Baltimore D. Activation in vitro of NF-kappa B by phosphorylation of its inhibitor I kappa B. Nature. 1990 Apr 12;344(6267):678–682. doi: 10.1038/344678a0. [DOI] [PubMed] [Google Scholar]
  26. Gorski K., Carneiro M., Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. doi: 10.1016/0092-8674(86)90519-2. [DOI] [PubMed] [Google Scholar]
  27. Hattori M., Abraham L. J., Northemann W., Fey G. H. Acute-phase reaction induces a specific complex between hepatic nuclear proteins and the interleukin 6 response element of the rat alpha 2-macroglobulin gene. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2364–2368. doi: 10.1073/pnas.87.6.2364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hocke G. M., Barry D., Fey G. H. Synergistic action of interleukin-6 and glucocorticoids is mediated by the interleukin-6 response element of the rat alpha 2 macroglobulin gene. Mol Cell Biol. 1992 May;12(5):2282–2294. doi: 10.1128/mcb.12.5.2282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Horellou P., Brundin P., Kalén P., Mallet J., Björklund A. In vivo release of dopa and dopamine from genetically engineered cells grafted to the denervated rat striatum. Neuron. 1990 Oct;5(4):393–402. doi: 10.1016/0896-6273(90)90078-t. [DOI] [PubMed] [Google Scholar]
  30. Huber P., Laurent M., Dalmon J. Human beta-fibrinogen gene expression. Upstream sequences involved in its tissue specific expression and its dexamethasone and interleukin 6 stimulation. J Biol Chem. 1990 Apr 5;265(10):5695–5701. [PubMed] [Google Scholar]
  31. Ito T., Tanahashi H., Misumi Y., Sakaki Y. Nuclear factors interacting with an interleukin-6 responsive element of rat alpha 2-macroglobulin gene. Nucleic Acids Res. 1989 Nov 25;17(22):9425–9435. doi: 10.1093/nar/17.22.9425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kessler D. S., Veals S. A., Fu X. Y., Levy D. E. Interferon-alpha regulates nuclear translocation and DNA-binding affinity of ISGF3, a multimeric transcriptional activator. Genes Dev. 1990 Oct;4(10):1753–1765. doi: 10.1101/gad.4.10.1753. [DOI] [PubMed] [Google Scholar]
  33. Kinoshita S., Akira S., Kishimoto T. A member of the C/EBP family, NF-IL6 beta, forms a heterodimer and transcriptionally synergizes with NF-IL6. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1473–1476. doi: 10.1073/pnas.89.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Krause E., Wegenka U., Möller C., Horn F., Heinrich P. C. Gene expression of the high molecular weight proteinase inhibitor alpha 2-macroglobulin. Biol Chem Hoppe Seyler. 1992 Jul;373(7):509–515. doi: 10.1515/bchm3.1992.373.2.509. [DOI] [PubMed] [Google Scholar]
  35. Kunz D., Zimmermann R., Heisig M., Heinrich P. C. Identification of the promoter sequences involved in the interleukin-6 dependent expression of the rat alpha 2-macroglobulin gene. Nucleic Acids Res. 1989 Feb 11;17(3):1121–1138. doi: 10.1093/nar/17.3.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  37. Lenardo M. J., Baltimore D. NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control. Cell. 1989 Jul 28;58(2):227–229. doi: 10.1016/0092-8674(89)90833-7. [DOI] [PubMed] [Google Scholar]
  38. Lord K. A., Abdollahi A., Thomas S. M., DeMarco M., Brugge J. S., Hoffman-Liebermann B., Liebermann D. A. Leukemia inhibitory factor and interleukin-6 trigger the same immediate early response, including tyrosine phosphorylation, upon induction of myeloid leukemia differentiation. Mol Cell Biol. 1991 Sep;11(9):4371–4379. doi: 10.1128/mcb.11.9.4371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Mann E. A., Croyle M. L., Lingrel J. B. Identification of sequences mediating interleukin-6 induction of a rat T kininogen gene. J Biol Chem. 1991 Sep 5;266(25):16931–16934. [PubMed] [Google Scholar]
  40. Marsh J. L., Erfle M., Wykes E. J. The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene. 1984 Dec;32(3):481–485. doi: 10.1016/0378-1119(84)90022-2. [DOI] [PubMed] [Google Scholar]
  41. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  42. Miskimins W. K., Roberts M. P., McClelland A., Ruddle F. H. Use of a protein-blotting procedure and a specific DNA probe to identify nuclear proteins that recognize the promoter region of the transferrin receptor gene. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6741–6744. doi: 10.1073/pnas.82.20.6741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Nakajima K., Wall R. Interleukin-6 signals activating junB and TIS11 gene transcription in a B-cell hybridoma. Mol Cell Biol. 1991 Mar;11(3):1409–1418. doi: 10.1128/mcb.11.3.1409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Natsuka S., Isshiki H., Akira S., Kishimoto T. Augmentation of haptoglobin production in Hep3B cell line by a nuclear factor NF-IL6. FEBS Lett. 1991 Oct 7;291(1):58–62. doi: 10.1016/0014-5793(91)81103-f. [DOI] [PubMed] [Google Scholar]
  45. Nonaka M., Huang Z. M. Interleukin-1-mediated enhancement of mouse factor B gene expression via NF kappa B-like hepatoma nuclear factor. Mol Cell Biol. 1990 Dec;10(12):6283–6289. doi: 10.1128/mcb.10.12.6283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Oliviero S., Cortese R. The human haptoglobin gene promoter: interleukin-6-responsive elements interact with a DNA-binding protein induced by interleukin-6. EMBO J. 1989 Apr;8(4):1145–1151. doi: 10.1002/j.1460-2075.1989.tb03485.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Osborn L., Kunkel S., Nabel G. J. Tumor necrosis factor alpha and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor kappa B. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2336–2340. doi: 10.1073/pnas.86.7.2336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Poli V., Cortese R. Interleukin 6 induces a liver-specific nuclear protein that binds to the promoter of acute-phase genes. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8202–8206. doi: 10.1073/pnas.86.21.8202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Poli V., Mancini F. P., Cortese R. IL-6DBP, a nuclear protein involved in interleukin-6 signal transduction, defines a new family of leucine zipper proteins related to C/EBP. Cell. 1990 Nov 2;63(3):643–653. doi: 10.1016/0092-8674(90)90459-r. [DOI] [PubMed] [Google Scholar]
  50. Rixon M. W., Chung D. W., Davie E. W. Nucleotide sequence of the gene for the gamma chain of human fibrinogen. Biochemistry. 1985 Apr 9;24(8):2077–2086. doi: 10.1021/bi00329a041. [DOI] [PubMed] [Google Scholar]
  51. Ron D., Brasier A. R., McGehee R. E., Jr, Habener J. F. Tumor necrosis factor-induced reversal of adipocytic phenotype of 3T3-L1 cells is preceded by a loss of nuclear CCAAT/enhancer binding protein (C/EBP). J Clin Invest. 1992 Jan;89(1):223–233. doi: 10.1172/JCI115566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Sarcione E. J., Bogden A. E. Hepatic synthesis of alpha 2 (acute phase)-globulin of rat plasma. Science. 1966 Jul 29;153(3735):547–548. doi: 10.1126/science.153.3735.547. [DOI] [PubMed] [Google Scholar]
  53. Sawadogo M., Van Dyke M. W., Gregor P. D., Roeder R. G. Multiple forms of the human gene-specific transcription factor USF. I. Complete purification and identification of USF from HeLa cell nuclei. J Biol Chem. 1988 Aug 25;263(24):11985–11993. [PubMed] [Google Scholar]
  54. Shapiro D. J., Sharp P. A., Wahli W. W., Keller M. J. A high-efficiency HeLa cell nuclear transcription extract. DNA. 1988 Jan-Feb;7(1):47–55. doi: 10.1089/dna.1988.7.47. [DOI] [PubMed] [Google Scholar]
  55. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  56. Tsuchiya Y., Hattori M., Hayashida K., Ishibashi H., Okubo H., Sakaki Y. Sequence analysis of the putative regulatory region of rat alpha 2-macroglobulin gene. Gene. 1987;57(1):73–80. doi: 10.1016/0378-1119(87)90178-8. [DOI] [PubMed] [Google Scholar]
  57. Wilson D. R., Juan T. S., Wilde M. D., Fey G. H., Darlington G. J. A 58-base-pair region of the human C3 gene confers synergistic inducibility by interleukin-1 and interleukin-6. Mol Cell Biol. 1990 Dec;10(12):6181–6191. doi: 10.1128/mcb.10.12.6181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Won K. A., Baumann H. The cytokine response element of the rat alpha 1-acid glycoprotein gene is a complex of several interacting regulatory sequences. Mol Cell Biol. 1990 Aug;10(8):3965–3978. doi: 10.1128/mcb.10.8.3965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Woods D. B., Ghysdael J., Owen M. J. Identification of nucleotide preferences in DNA sequences recognised specifically by c-Ets-1 protein. Nucleic Acids Res. 1992 Feb 25;20(4):699–704. doi: 10.1093/nar/20.4.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Zhang Y. H., Lin J. X., Vilcek J. Interleukin-6 induction by tumor necrosis factor and interleukin-1 in human fibroblasts involves activation of a nuclear factor binding to a kappa B-like sequence. Mol Cell Biol. 1990 Jul;10(7):3818–3823. doi: 10.1128/mcb.10.7.3818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Zohlnhöfer D., Graeve L., Rose-John S., Schooltink H., Dittrich E., Heinrich P. C. The hepatic interleukin-6 receptor. Down-regulation of the interleukin-6 binding subunit (gp80) by its ligand. FEBS Lett. 1992 Jul 20;306(2-3):219–222. doi: 10.1016/0014-5793(92)81004-6. [DOI] [PubMed] [Google Scholar]

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