Critical role of the automodification of poly(ADP-ribose) polymerase-1 in nuclear factor-kappaB-dependent gene expression in primary cultured mouse glial cells
| Autor: | Hiroshi Nagaso, Hidemitsu Nakajima, Shigeru Hoshiko, Midori Ishikawa, Nobukazu Kakui, Toyokazu Hiranuma |
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| Rok vydání: | 2004 |
| Předmět: |
Lipopolysaccharides
Transcriptional Activation Time Factors Transcription Genetic Cell Survival Poly ADP ribose polymerase Blotting Western Nitric Oxide Synthase Type II Biology Biochemistry Mice Transcription (biology) Gene expression Transcriptional regulation Animals Immunoprecipitation Electrophoretic mobility shift assay RNA Antisense RNA Messenger Molecular Biology Cells Cultured Nitrites Regulation of gene expression Cell Nucleus Inflammation Messenger RNA Mice Inbred BALB C Dose-Response Relationship Drug Oligonucleotide Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha NF-kappa B NF-kappa B p50 Subunit Cell Biology DNA Oligonucleotides Antisense Molecular biology Up-Regulation Protein Transport Gene Expression Regulation Cytokines Nitric Oxide Synthase Poly(ADP-ribose) Polymerases Neuroglia Transcription Factors |
| Zdroj: | The Journal of biological chemistry. 279(41) |
| ISSN: | 0021-9258 |
| Popis: | Synthesis of ADP-ribose polymers catalyzed by poly-(ADP-ribose) polymerase-1 (PARP-1) has been implicated in transcriptional regulation. Recent studies with PARP-1 null mice and PARP-1 inhibitors have also demonstrated that PARP-1 has an essential role in nuclear factor-kappaB (NF-kappaB)-dependent gene expression induced by various inflammatory stimuli. In this study, we used primary cultured mouse glial cells to investigate the role of poly(ADP-ribosyl)ation by PARP-1 in NF-kappaB-dependent gene expression. PARP-1 inhibitors and the antisense RNA for PARP-1 mRNA suppressed lipopolysaccharide (LPS)-induced expression of tumor necrosis factor-alpha and inducible nitric-oxide synthase, suggesting that PARP-1 activity has a critical role in synthesis. Western blotting with anti-poly(ADP-ribose) antibody revealed that PARP-1 itself was mainly poly(ADP-ribosyl)ated in glial cells, i.e. automodified PARP-1 (AM-PARP). The amounts of AM-PARP were not affected by LPS treatment, but were decreased by PARP-1 inhibitors. Electrophoretic mobility shift assay revealed that PARP-1 inhibitors and the antisense RNA for PARP-1 mRNA reduced the LPS-induced DNA binding of NF-kappaB. Non-modified PARP-1 also reduced the DNA binding of NF-kappaB via its physical association with NF-kappaB, whereas AM-PARP had no effect. On the other hand, enhancement of the automodification of PARP-1 by the addition of NAD+, its substrate, promoted the DNA binding of NF-kappaB. Furthermore, in in vitro transcription assay, the addition of AM-PARP or NAD+ to nuclear extracts promoted NF-kappaB p50-dependent transcription. These results indicate that automodification of PARP-1 positively up-regulates formation of the NF-kappaB.DNA complex and enhances transcriptional activation. Therefore, AM-PARP may be critical for the NF-kappaB-dependent gene expression of some inflammatory mediators in glial cells. |
| Databáze: | OpenAIRE |
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