Characterization of the promoter for the human long pentraxin PTX3 - Role of NF-kappa B in tumor necrosis factor-alpha and interleukin-1 beta regulation

The ''long pentraxins'' are an emerging family of genes that have conserved in their carboxy-terminal halves a pentraxin domain homologous to the prototypical acute phase protein pentraxins (C-reactive protein and serum amyloid P component) and acquired novel amino-terminal domains. In this report, a genomic fragment of 1371 nucleotides from the human ''long pentraxin'' gene PTX3 is characterized as a promoter on tumor necrosis factor-alpha (TNF alpha) and interleukin (IL)-1 beta exposure in transfected 8387 human fibroblasts by chloramphenicol acetyltransferase and RNase protection assays. In the same cells, the PTX3 promoter does not respond to IL-6 stimulation. Furthermore, IL-1 beta and TNF alpha responsiveness is not seen in the Hep 3B hepatoma cell line, The minimal promoter contains one NF-kappa B element which is shown to be necessary for induction and able to bind p50 homodimers and p65 heterodimers but not c-Rel. Mutants in this site lose the ability to bind NF-kappa B proteins and to respond to TNF alpha and IL-1 beta in functional assays. Sp1- and AP-1 binding sites lying in proximity to the NF-kappa B site do not seem to play a major role for cytokine responsiveness. Finally, cotransfection experiments with expression vectors validate that the natural promoter contains a functional NF-kappa B site.