The protein product of the early response gene, Egr-1, is required for macrophage differentiation. Egr-1 mRNA is induced by lipopolysaccharide within minutes in murine peritoneal macrophages through an effect on the rate of transcription of Egr-1. Therefore, we undertook a series of studies to characterize Egr-1 protein induced by LPS and to define enhancer elements in the Egr-1 promoter which mediate the effect of LPS in murine macrophages. Salmonella minnesota Re 595 LPS (10 ng/ml or higher) induced Egr-1 protein of 80 kDa within 45 min in thioglycollate-elicited peritoneal macrophages. The promoter region of Egr-1 contains six serum response elements (SRE) which each contain a core CArG box. CAT activity in LPStreated macrophages transfected with Egr-1-CAT constructs containing the two most upstream 5' SREs was increased 4.7-fold whereas CAT activity in macrophages transfected with constructs lacking these two most upstream SREs was not affected by LPS. LPS induced CAT activity in macrophages transfected with constructs containing tetramers of each of the two most 5' SREs upstream of a heterologous promoter linked to CAT. Mutation of the CArG box abolished the effect of LPS. These findings suggest that the CArG box elements within the two most upstream SREs may mediate the effect of LPS on transcription of Egr-1.
Get full access to this article
View all access options for this article.
References
1.
Sukhatme VP, Cao X., Chang LL et al. A zinc-finger encoding gene coregulated with c-Fos during growth and differentiation and after depolarization. Cell1988; 53: 37-43.
2.
Sukhatme VPEarly transcriptional events in cell growth: the Egr-1 family. J Am Soc Nephrol1990; 1: 859-866.
3.
Cao X., Koski RA, Gashler A. et al. Identification and characterization of the Egr-1 gene product, a DNA-binding zinc finger protein induced by differentiation and growth signals . Mol Cell Biol1990; 10: 1931-1939.
4.
Pavletich NP, Pabo COZinc finger-DNA recognition: crystal structure of a zif 268-DNA complex at 2.1 Å. Science1991; 252: 809-817.
5.
Lemaire P., Vesque C., Schmitt J., Stunnenberg H., Frank R., Charnay P.The serum-inducible mouse gene Krox-24 encodes a sequence-specific transcriptional activator. Mol Cell Biol1990; 10: 3456-3467.
6.
Nguyen HQ, Hoffman-Liebermann B., Liebermann DAThe zinc finger transcription factor Egr-1 is essential for and restricts differentiation along the macrophage lineage. Cell1993; 72: 197-209.
7.
Coleman DL, Bartiss A., Liu J., Sukhatme VP, Rupprecht HDLipopolysaccharide induces Egr-1 mRNA and protein in murine peritioneal macrophages . J Immunol1992; 149: 3045-3051.
8.
Liu J., Lacy J., Sukhatme VP, Coleman DLGranulocyte-macrophage colony-stimulating factor induces transcriptional activation of Egr-1 in murine peritoneal macrophages. J Biol Chem1991; 266: 5929-5933.
9.
Raetz Crh, Ulevitch RJ, Wright SD, Sibley CH, Ding A., Nathan CFGram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J1991; 5: 2652-2660.
10.
Takayama K. , Mitchell DH, Din AA, Mukerjee P., Li C., Coleman DLMonomeric Re lipopolysaccharide from Escherichia coli is more active than the aggregated form in the Limulus amebocyte lysate assay and in inducing Egr-1 mRNA in murine peritoneal macrophages. J Biol Chem1994; 269: 2241-2244.
11.
Tsai-Morris C-H., Cao X., Sukhatme VP5' flanking sequence and genomic structure of Egr-1, a murine mitogen inducible zinc finger encoding gene. Nucleic Acids Res1988; 16: 8835-8846.
12.
Christy BA, Nathans D.Functional serum response elements upstream of the growth factor inducible gene zif 268. Mol Cell Biol1989; 9: 4889-4895.
13.
Rupprecht HD , Sukhatme VP, Lacy J., Sterzel RB, Coleman DLPDGF-induced Egr-1 expression in rat mesangial cells is mediated through upstream serum response elements. Am J Physiology1992; 263: F623-F636.
14.
Rupprecht HD , Sukhatme VP, Rupprecht AP, Sterzel RB, Coleman DLSerum response elements mediate protein kinase C dependent transcriptional induction of early growth response gene-1 by arginine vasopressin in rat mesangial cells. J Cell Physiol1994; 159: 311-323.
15.
Gius D., Cao X., Rauscher III FJ, Cohen DR, Curran T., Sukhatme VPTranscriptional activation and repression by Fos are independent functions: the C-terminus represses immediate-early gene expression via CArG elements . Mol Cell Biol1990; 10: 4243-4255.
16.
Muller JM, Ziegler-Heitbrock HW, Baeuerle PANuclear factor kappa B, a mediator of lipopolysaccharide effects. Immunobiology1993; 187: 233-256.
17.
Ohmori Y., Hamilton TACooperative interaction between interferon (IFN) stimulus response element and Kappa B sequence motifs controls IFN gamma and lipopolysaccharide-stimulated transcription from the murine IP-10 promoter. J Biol Chem1993; 268: 6677-6688.
18.
Kaminska B. , Kaczmarek L., Malaguarnera L. et al. Transcription factor activation and functional stimulation of human monocytes. Cell Biol Intern Rep1992; 16: 37-45.
19.
Zhang Y., Rom WNRegulation of the interleukin-1 beta gene by mycobacterial components and lipopolysaccharide is medicated by two nuclear factor-IL6 motifs. Mol Cell Biol1993; 13: 3831-3837.
20.
Bernstein S.H. , Kharbanda S.M., Sherman M.L., Sukhatme V.P., Kufe DWPostranscriptional regulation of the zinc finger-encoding Egr-1 gene by granulocyte-macrophage colony stimulating factor in human U937 monocytic leukemia cells: Involvement of a pertussis toxin-sensitive G protein. Cell Growth Diff1991; 2: 272-278.
21.
Rupprecht AP , Coleman DLTransfection of adherent murine peritoneal macrophages with a reporter gene using DEAE-dextran. J Immunol Meth1991; 144: 157-163.
22.
Herrera RE, Shaw PE, Nordheim A.Occupation of the c-fos serum response element in vivo by a multi-protein complex is unaltered by growth factor induction. Nature1989; 340: 68-70.
23.
Datta R., Taneja N., Sukhatme VP, Qureshi SA, Weichselbaum R., Kufe DWReactive oxygen intermediates target CC(A/T)6GG sequences to mediate activation of the early growth response 1 transcription factor gene by ionizing radiation. Proc Natl Acad Sci USA1993; 90: 2419-2422.
24.
Alexandropoulos K., Qureshi SA, Rim M., Sukhatme VP, Foster DAv-Fps-responsiveness in the Egr-1 promoter is mediated by serum response elements. Nucleic Acids Res1992; 20: 2355-2359.
25.
Norman C., Runswick M, Pollock R, Treisman R. Isolation and properties of cDNA clones encoding SRF, a transcription factor that binds to the c-fos serum response element. Cell1988; 55: 989-1003.
26.
Shaw PE, Schroeter H., Nordheim A.The ability of a ternary complex to form over the serum response element correlates with serum inducibility of the human c-fos promoter. Cell1989; 56: 563-572.
27.
Treisman R.Identification and purification of a polypeptide that binds to the c-fos serum response element. EMBO J1987; 6: 2711-2717.
28.
Attar RM, Gilman MZExpression cloning of a novel zinc finger protein that binds to the c-fos serum response element. Mol Cell Biol1992 ; 12: 2432-2443.
29.
Donovan-Peluso M., George LD, Hassett ACLipopolysaccharide induction of tissue factor expression in THP-1 monocytic cells: protein-DNA interactions with the promoter . J Biol Chem1994; 269: 1361-1369.
30.
Ulich TR, Guo KZ, Irwin B., Remick DG, Davatelis GNEndotoxin-induced cytokine gene expression in vivo. II: Regulation of tumor necrosis factor and interleukin-1 alpha/beta expression and suppression. Am J Pathol1990; 137: 1173-1185.
31.
Shakhov AN, Collart MA, Vassalli P., Nedospasov SA, Jongeneel CVKappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor alpha gene in primary macrophages . J Exp Med1990; 171: 35-47.
32.
Engstrom Y. , Kadalayil L., Sun SC, Samakovlis S., Hultmark K., Faye I.Kappa B-like motifs regulate the induction of immune genes in Drosophila. J Mol Biol1993; 232: 327-333.
33.
Kappler C., Meister M., Lagueux M., Gateff E., Hoffman JA, Reichhart JMInsect immunity: two 17 bp repeats nesting a kappa B-related sequence confer inducibility to the diptericin gene and bind a polypeptide in bacteria-challenged Drosophila. EMBO J1993; 12: 1561-1568.
34.
Grove M., Plumb M.C/EBP, NF-kappa B, and c-Ets family members and transcriptional regulation of the cell-specific and inducible macrophage inflammatory protein 1 alpha immediate-early gene. Mol Cell Biol1993; 13: 5276-5289.
Collart MA, Baeuerle P., Vassalli P.Regulation of tumor necrosis factor alpha transcription in macrophages: Involvement of four kappa B-like motifs and of constitutive and inducible forms of NF-kappa B. Mol Cell Biol1990; 10: 1498-1506.
37.
Shirakawa F. , Laito K., Bonagura CA et al. The human prointerleukin 1 beta gene requires DNA sequences both proximal and distal to the transcription start site for tissue-specific induction. Mol Cell Biol1993; 13: 1332-1344.
