Title | Poly(ADP-ribose) polymerase-1-induced NAD(+) depletion promotes nuclear factor-κB transcriptional activity by preventing p65 de-acetylation. |
Publication Type | Journal Article |
Year of Publication | 2013 |
Authors | Kauppinen TM, Gan L, Swanson RA |
Journal | Biochim Biophys Acta |
Volume | 1833 |
Issue | 8 |
Pagination | 1985-91 |
Date Published | 2013 Aug |
ISSN | 0006-3002 |
Keywords | Acetylation, Animals, Astrocytes, Carbazoles, Cells, Cultured, eIF-2 Kinase, Humans, Methylnitronitrosoguanidine, Mice, NAD, NF-kappa B, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerase Inhibitors, Poly(ADP-ribose) Polymerases, Sirtuin 1, Transcription, Genetic, Transcriptional Activation, Transfection |
Abstract | NF-κB is a transcription factor that integrates pro-inflammatory and pro-survival responses in diverse cell types. The activity of NF-κB is regulated in part by acetylation of its p65 subunit at lysine 310, which is required for transcription complex formation. De-acetylation at this site is performed by sirtuin 1(SIRT1) and possibly other sirtuins in an NAD(+) dependent manner, such that SIRT1 inhibition promotes NF-κB transcriptional activity. It is unknown, however, whether changes in NAD(+) levels can influence p65 acetylation and cellular inflammatory responses. Poly(ADP-ribose)-1 (PARP-1) is an abundant nuclear enzyme that consumes NAD(+) in the process of forming (ADP-ribose)polymers on target proteins, and extensive PARP-1 activation can reduce intracellular NAD(+) concentrations. Here we tested the idea that PARP-1 activation can regulate NF-κB transcriptional activity by reducing NAD(+) concentrations and thereby inhibiting de-acetylation of p65. Primary astrocyte cultures were treated with the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) to induce PARP-1 activation. This resulted in sustained acetylation of p65 and increased NF-κB transcriptional activity as monitored by a κB-driven eGFP reporter gene. These effects of MNNG were negated by a PARP-1 inhibitor, in PARP-1(-/-) cells, and in PARP-1(-/-) cells transfected with a catalytically inactive PARP-1 construct, thus confirming that these effects are mediated by PARP-1 catalytic activity. The effects of PARP-1 activation were replicated by a SIRT1 inhibitor, EX-527, and were reversed by exogenous NAD(+). These findings demonstrate that PARP-1-induced changes in NAD(+) levels can modulate NF-κB transcriptional activity through effects on p65 acetylation. |
DOI | 10.1016/j.bbamcr.2013.04.005 |
Alternate Journal | Biochim. Biophys. Acta |
PubMed ID | 23597856 |
PubMed Central ID | PMC4041949 |
Grant List | P50 AG023501 / AG / NIA NIH HHS / United States R01 AG036884 / AG / NIA NIH HHS / United States R01 NS041421 / NS / NINDS NIH HHS / United States P01 AG019724 / AG / NIA NIH HHS / United States R01 AG033017 / AG / NIA NIH HHS / United States |