Characterization of the promoter for the human long pentraxin PTX3. Role of NF-kappaB in tumor necrosis factor-alpha and interleukin-1beta 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 (TNFalpha) and interleukin (IL)-1beta 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-1beta and TNFalpha responsiveness is not seen in the Hep 3B hepatoma cell line. The minimal promoter contains one NF-kappaB 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-kappaB proteins and to respond to TNFalpha and IL-1beta in functional assays. Sp1- and AP-1 binding sites lying in proximity to the NF-kappaB 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-kappaB site.
Basile
A
A
Istituto di Ricerche Farmacologiche "Mario Negri," via Eritrea, 62, 20157 Milan, Italy.
Sica
A
A
d'Aniello
E
E
Breviario
F
F
Garrido
G
G
Castellano
M
M
Mantovani
A
A
Introna
M
M
D000818:Animals; D001483:Base Sequence; D002097:C-Reactive Protein / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012260:Ribonucleases / Q000378:metabolism; D016415:Sequence Alignment; D017384:Sequence Deletion; D000683:Serum Amyloid P-Component / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D000099153:Pentraxins
D002097:C-Reactive Protein; D007375:Interleukin-1; D016328:NF-kappa B; D012260:Ribonucleases; D000683:Serum Amyloid P-Component; D014409:Tumor Necrosis Factor-alpha; D000099153:Pentraxins