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. 1994 Sep-Dec;67(5-6):269–276.

Analysis of Egr-1 protein induction in murine peritoneal macrophages treated with granulocyte-macrophage colony-stimulating factor.

C Li 1, D H Mitchell 1, D L Coleman 1
PMCID: PMC2588905  PMID: 8579643

Abstract

Transcription of early growth response gene-1 (Egr-1) is required for macrophage differentiation. Since granulocyte-macrophage colony-stimulating factor (GM-CSF) induces macrophage proliferation as well as the transcription of Egr-1 mRNA, we characterized the induction of Egr-1 protein in murine peritoneal macrophages following treatment with GM-CSF. As determined by Western blot analysis, the concentration of GM-CSF required to induce detectable Egr-1 protein in macrophages was 50 units/ml, and the inducible Egr-1 protein species was 80 kDa. Following stimulation of macrophages with GM-CSF, Egr-1 protein was detected within 25 min and reached maximum level at 70 min. The concentration of GM-CSF that was required to induce Egr-1 protein in macrophages was similar to that required to induce macrophage proliferation. A similar concentration has been detected previously in mouse serum exposed to bacterial endotoxin in vivo, suggesting that the 80 kDa Egr-1 protein may be induced in macrophages under the physiologic circumstances of pathogen invasion. Thus, our data support further study on the role of Egr-1 protein in mediating GM-CSF induction of macrophage proliferation.

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Selected References

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

  1. Alexandropoulos K., Qureshi S. A., Rim M., Sukhatme V. P., Foster D. A. v-Fps-responsiveness in the Egr-1 promoter is mediated by serum response elements. Nucleic Acids Res. 1992 May 11;20(9):2355–2359. doi: 10.1093/nar/20.9.2355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bernstein S. H., Kharbanda S. M., Sherman M. L., Sukhatme V. P., Kufe D. W. Posttranscriptional regulation of the zinc finger-encoding EGR-1 gene by granulocyte-macrophage colony-stimulating factor in human U-937 monocytic leukemia cells: involvement of a pertussis toxin-sensitive G protein. Cell Growth Differ. 1991 Jun;2(6):273–278. [PubMed] [Google Scholar]
  3. Cao X. M., Koski R. A., Gashler A., McKiernan M., Morris C. F., Gaffney R., Hay R. V., Sukhatme V. P. Identification and characterization of the Egr-1 gene product, a DNA-binding zinc finger protein induced by differentiation and growth signals. Mol Cell Biol. 1990 May;10(5):1931–1939. doi: 10.1128/mcb.10.5.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cao X., Mahendran R., Guy G. R., Tan Y. H. Protein phosphatase inhibitors induce the sustained expression of the Egr-1 gene and the hyperphosphorylation of its gene product. J Biol Chem. 1992 Jun 25;267(18):12991–12997. [PubMed] [Google Scholar]
  5. Clark S. C., Kamen R. The human hematopoietic colony-stimulating factors. Science. 1987 Jun 5;236(4806):1229–1237. doi: 10.1126/science.3296190. [DOI] [PubMed] [Google Scholar]
  6. Coleman D. L., Liu J., Bartiss A. H. Recombinant granulocyte-macrophage colony-stimulating factor increases adenylate cyclase activity in murine peritoneal macrophages. J Immunol. 1989 Dec 15;143(12):4134–4140. [PubMed] [Google Scholar]
  7. Hazel T. G., Nathans D., Lau L. F. A gene inducible by serum growth factors encodes a member of the steroid and thyroid hormone receptor superfamily. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8444–8448. doi: 10.1073/pnas.85.22.8444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Iwaki K., Sukhatme V. P., Shubeita H. E., Chien K. R. Alpha- and beta-adrenergic stimulation induces distinct patterns of immediate early gene expression in neonatal rat myocardial cells. fos/jun expression is associated with sarcomere assembly; Egr-1 induction is primarily an alpha 1-mediated response. J Biol Chem. 1990 Aug 15;265(23):13809–13817. [PubMed] [Google Scholar]
  9. Joseph L. J., Le Beau M. M., Jamieson G. A., Jr, Acharya S., Shows T. B., Rowley J. D., Sukhatme V. P. Molecular cloning, sequencing, and mapping of EGR2, a human early growth response gene encoding a protein with "zinc-binding finger" structure. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7164–7168. doi: 10.1073/pnas.85.19.7164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kharbanda S., Rubin E., Datta R., Hass R., Sukhatme V., Kufe D. Transcriptional regulation of the early growth response 1 gene in human myeloid leukemia cells by okadaic acid. Cell Growth Differ. 1993 Jan;4(1):17–23. [PubMed] [Google Scholar]
  11. 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]
  12. Liu J. W., Lacy J., Sukhatme V. P., Coleman D. L. Granulocyte-macrophage colony-stimulating factor induces transcriptional activation of Egr-1 in murine peritoneal macrophages. J Biol Chem. 1991 Mar 25;266(9):5929–5933. [PubMed] [Google Scholar]
  13. Madden S. L., Cook D. M., Morris J. F., Gashler A., Sukhatme V. P., Rauscher F. J., 3rd Transcriptional repression mediated by the WT1 Wilms tumor gene product. Science. 1991 Sep 27;253(5027):1550–1553. doi: 10.1126/science.1654597. [DOI] [PubMed] [Google Scholar]
  14. Nguyen H. Q., Hoffman-Liebermann B., Liebermann D. A. The zinc finger transcription factor Egr-1 is essential for and restricts differentiation along the macrophage lineage. Cell. 1993 Jan 29;72(2):197–209. doi: 10.1016/0092-8674(93)90660-i. [DOI] [PubMed] [Google Scholar]
  15. Nielsen D. A., Chou J., MacKrell A. J., Casadaban M. J., Steiner D. F. Expression of a preproinsulin-beta-galactosidase gene fusion in mammalian cells. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5198–5202. doi: 10.1073/pnas.80.17.5198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ouellette A. J., Malt R. A., Sukhatme V. P., Bonventre J. V. Expression of two "immediate early" genes, Egr-1 and c-fos, in response to renal ischemia and during compensatory renal hypertrophy in mice. J Clin Invest. 1990 Mar;85(3):766–771. doi: 10.1172/JCI114502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Qureshi S. A., Cao X. M., Sukhatme V. P., Foster D. A. v-Src activates mitogen-responsive transcription factor Egr-1 via serum response elements. J Biol Chem. 1991 Jun 15;266(17):10802–10806. [PubMed] [Google Scholar]
  18. Ragona G., Edwards S. A., Mercola D. A., Adamson E. D., Calogero A. The transcriptional factor Egr-1 is synthesized by baculovirus-infected insect cells in an active, DNA-binding form. DNA Cell Biol. 1991 Jan-Feb;10(1):61–66. doi: 10.1089/dna.1991.10.61. [DOI] [PubMed] [Google Scholar]
  19. Ruef C., Coleman D. L. Granulocyte-macrophage colony-stimulating factor: pleiotropic cytokine with potential clinical usefulness. Rev Infect Dis. 1990 Jan-Feb;12(1):41–62. doi: 10.1093/clinids/12.1.41. [DOI] [PubMed] [Google Scholar]
  20. Rupprecht H. D., Dann P., Sukhatme V. P., Sterzel R. B., Coleman D. L. Effect of vasoactive agents on induction of Egr-1 in rat mesangial cells: correlation with mitogenicity. Am J Physiol. 1992 Oct;263(4 Pt 2):F623–F636. doi: 10.1152/ajprenal.1992.263.4.F623. [DOI] [PubMed] [Google Scholar]
  21. Rupprecht H. D., Sukhatme V. P., Lacy J., Sterzel R. B., Coleman D. L. PDGF-induced Egr-1 expression in rat mesangial cells is mediated through upstream serum response elements. Am J Physiol. 1993 Sep;265(3 Pt 2):F351–F360. doi: 10.1152/ajprenal.1993.265.3.F351. [DOI] [PubMed] [Google Scholar]
  22. Ryder K., Lanahan A., Perez-Albuerne E., Nathans D. jun-D: a third member of the jun gene family. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1500–1503. doi: 10.1073/pnas.86.5.1500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Seyfert V. L., McMahon S., Glenn W., Cao X. M., Sukhatme V. P., Monroe J. G. Egr-1 expression in surface Ig-mediated B cell activation. Kinetics and association with protein kinase C activation. J Immunol. 1990 Dec 1;145(11):3647–3653. [PubMed] [Google Scholar]
  24. Sukhatme V. P. Early transcriptional events in cell growth: the Egr family. J Am Soc Nephrol. 1990 Dec;1(6):859–866. doi: 10.1681/ASN.V16859. [DOI] [PubMed] [Google Scholar]
  25. Suva L. J., Ernst M., Rodan G. A. Retinoic acid increases zif268 early gene expression in rat preosteoblastic cells. Mol Cell Biol. 1991 May;11(5):2503–2510. doi: 10.1128/mcb.11.5.2503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Waters C. M., Hancock D. C., Evan G. I. Identification and characterisation of the egr-1 gene product as an inducible, short-lived, nuclear phosphoprotein. Oncogene. 1990 May;5(5):669–674. [PubMed] [Google Scholar]
  27. Zerial M., Toschi L., Ryseck R. P., Schuermann M., Müller R., Bravo R. The product of a novel growth factor activated gene, fos B, interacts with JUN proteins enhancing their DNA binding activity. EMBO J. 1989 Mar;8(3):805–813. doi: 10.1002/j.1460-2075.1989.tb03441.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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