Evidence for regulation of NF-κB by poly(ADP-ribose) polymerase

Abstract

The DNA-binding activity of NF-kappaB in nuclear extracts of poly(ADP-ribose) polymerase (PARP)-defective mutant L1210 cell clones was markedly increased and was inversely correlated with the PARP content in these cells. The DNA-binding activity of NF-kappaB in a clone with the lowest PARP content (Cl-3527, contained 6% of PARP of wild type cells) was about 35-fold of that of the wild-type cells, whereas the change in the DNA-binding activity of AP-1 and SP-1 in the mutant was relatively small or not so significant. Transfection of a PARP-expressing plasmid to the mutant cells decreased the abnormally high levels of NF-kappaB complexes, especially p50/p65(Rel A) complex, to near the normal level. Moreover, poly(ADP-ribosyl)ation of nuclear extracts in vitro suppressed the ability of NF-kappaB to form a complex with its specific DNA probe by approx. 80%. Further analysis with purified recombinant NF-kappaB proteins revealed that both rp50 and rMBP-p65 (Rel A) proteins, but not rGST-IkappaB, could be poly(ADP-ribosyl)ated in vitro and that the modification resulted in a marked decrease in the DNA-binding activity of rMBP-p65, whereas a slight activation was observed in rp50. Poly(ADP-ribosyl)ated p65/NF-kappaB was detected in the cytosol of wild type L1210 cells by immunoblotting with anti-poly(ADP-ribose) and anti-p65 antibodies. Taken together, these results strongly suggest that PARP is involved in the regulation of NF-kappaB through the protein modification.

Poly(ADP-ribose) polymerase-1 is required for efficient HIV-1 integration

“…Transcription of NF-B is markedly reduced in PARP-1 Ϫ/Ϫ fibroblasts (43,44). PARP-1 could increase HIV-1 transcription by facilitating actions of NF-B (45,46). Additional evidence suggesting a role of PARP-1 in transcription comes from studies demonstrating a coactivator role for PARP-1 in regulating transcription of the HTLV-1 TAX protein (47).…”

Differential regulation of CXC ligand 1 transcription in melanoma cell lines by poly(ADP-ribose) polymerase-1

“…Whereas the glial cells showed diminished p38-mitogen-activated protein kinase activation as well as NF-kB DNA-binding and target gene expression, PARP-1 À/À macrophages only lacked in NF-kB activation. Furthermore, murine lymphocytic leukemia cells deficient in PARP-1 exhibited increased DNA-binding activity of NF-kB and transfection of these cells with a PARPexpressing plasmid decreased the high level of binding to normal levels (Kameoka et al, 2000). Inhibition of PARP-1 activity in airway epithelial cells, on the other hand, showed reduced NF-kB activation and reduced CXCL8 expression upon H 2 O 2 induction and prevented lung inflammation in vivo (Boulares et al, 2003).…”

Tumor necrosis factor signaling

“…[108][109][110] However, there are contradictory data concerning the importance of the PARP-1 enzymatic activity for NF-kB activation. 108,109,[111][112][113] TNF is able to activate PARP-1 via the production of ROS, which in turn causes PARP-inducing DNA damage. However, it is an open question whether TNF-induced PARP-1 activation via ROS is a prerequisite for its role in TNF-induced NF-kB activation or whether PARP-1 exerts its NF-kB-supporting capability independent of prior activation.…”