THE HUMAN BETA-FIBRINOGEN PROMOTER CONTAINS A HEPATOCYTE NUCLEAR FACTOR 1-DEPENDENT INTERLEUKIN-6-RESPONSIVE ELEMENT

Acute-phase reactants are liver proteins whose synthesis is positively or negatively regulated during inflammation. The main mediators of this phenomenon are glucocorticoids and interleukin-6 (IL-6), a pleiotropic cytokine that also controls hematopoiesis. Functional analysis of several acute-phase reactant promoter regions has identified two major DNA motifs used by IL-6-regulated genes. The first one corresponds to a CTGG(G/A)AA sequence, and the other is a binding site for members of the C/EBP family of nuclear proteins. We have previously shown that the human beta fibrinogen (betaFg) promoter contains an IL-6-responsive region, located between bp -150 and -67 (P. Huber, M. laurent, and J. Dalmon, J. Biol. Chem. 265:5695-5701, 1990). In this study, using DNase I footprinting, mobility shift assays, and mutagenesis, we demonstrate that at least three subdomains of this region are necessary to observe a full response to IL-6. The most distal contains a CTGGGAA motif, and its mutation inhibits IL-6 stimulation. Another, which is able to interact with several distinct nuclear proteins, among them members of the C/EBP family, is dispensable for IL-6 induction but plays an important role in the constitutive expression of betaFg. Finally, a proximal hepatocyte nuclear factor 1 binding site, already described as the major determinant of betaFg tissue-specific expression, is also required for IL-6 stimulation. These results indicate a complex interplay between nuclear proteins within the betaFg IL-6-responsive region and suggest a tight functional coupling between the tissue-specific and inducible elements.