Journal of Endotoxin Research

 

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Journal of Endotoxin Research, Vol. 2, No. 4, 239-246 (1995)
DOI: 10.1177/096805199500200403

Lipopolysaccharide induces expression of Egr-1 in macrophages through upstream CArG box (serum response) elements

C. Li

Infectious Disease Section, VA Medical Center, Yale University School of Medicine, West Haven, Connecticut and Infectious Disease Section, Department of Veterans Affairs Medical Center, Denver, Colorado, USA

D.H. Mitchell

Infectious Disease Section, VA Medical Center, Yale University School of Medicine, West Haven, Connecticut and Infectious Disease Section, Department of Veterans Affairs Medical Center, Denver, Colorado, USA

D.L. Coleman

Infectious Disease Section, VA Medical Center, Yale University School of Medicine, West Haven, Connecticut and Infectious Disease Section, Department of Veterans Affairs Medical Center, Denver, Colorado, USA

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.


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