Translational induction of liver-enriched transcriptional inhibitory protein during acute phase response leads to repression of CCAAT/enhancer binding protein alpha mRNA

Abstract

Lipopolysacharide (LPS) induced acute phase response (APR) in mouse liver leads to elevation of the low molecular weight CCAAT/Enhancer binding protein (C/EBP) beta isoform, liver-enriched transcriptional inhibitory protein (LIP). In this paper, we investigate the pathway for LIP induction during APR and the role of LIP in regulation of the C/EBPalpha promoter. The 5' region of C/EBPbeta mRNA has been shown to be involved in the regulation of LIP translation. Our data demonstrate that binding of cytoplasmic proteins to the 5' region of C/EBPbeta mRNA is altered in response to LPS administration. One of the major changes is induced binding of a cytoplasmic protein that is immunologically identical to the previously characterized RNA-binding protein CUGBP1. Induction of CUGBP1 binding activity in liver cytoplasm during APR is accompanied by the elevation of CUGBP1 binding activity on polysomes. CUGBP1 immunoprecipitated from livers of LPS-treated mice, but not from normal animals, is capable of inducing LIP translation in a cell-free translation system. The ability of CUGBP1 to induce LIP translation during APR depends on phosphorylation of CUGBP1. We show that elevation of LIP during APR and after partial hepatectomy leads to increased binding of LIP to the C/EBP consensus site found within the mouse C/EBPalpha promoter. This binding correlates with reduction of C/EBPalpha mRNA levels in both biological situations. Co-transfection experiments showed that full-length C/EBPbeta activates the C/EBPalpha promoter, while LIP blocks this activation. Our data suggest that the dominant negative isoform of C/EBPbeta, LIP, down-regulates the C/EBPalpha promoter in liver and in cultured hepatocytes. Because full-length C/EBPalpha and C/EBPbeta proteins regulate liver proliferation, this function of LIP may be important in liver growth and differentiation.