Poly ADP-Ribosylation Of Of HMGB1Enhances Acetylation

ObjectiveHMGB1is thought to be a key late-acting mediator of inflammation. HMGB1can be modified as acetylation, phosphorylation, methylation and ADP-ribosylation. Studies have shown that acetylation in HMGB1enhances the process of translocation from the nucleus to the cytoplasm and releasing into the extracellular space.In recent years, it is reported that PARP-1is involved in the process of intracellular localization and exocytosis of HMGB1, which may have relation with the poly ADP-ribosylation modification.This research studies the influence of poly ADP-ribosylation of HMGB1on acetylation, to find a novel mechanism by which PARP-1promotes the inflammation and provide more theoretical basis for the use of PARP-1inhibitors in the treatment of HMGB1-related inflammation.MethodsRAW264.7cells were treated with LPS(100ng/ml)or LPS+3-ABC10mmol/l) for2h and4h, protein is extracted with RIPA lysis; use immunoprecipitation(IP) with anti-HMGB1antibody in to enrich HMGB1; detect the poly ADP-ribosylation and acetylation of HMGB1by Western-blot with anti-PAR antibody and acet yl-lysine antibody. In vitro PARP-1modified HMGB1with6-biotin-17-NAD, proteins modified by biotin-NAD were detected by Western-blot with HRP-labeled streptavidin antibody; in vitro CBP acetylated HMGB1, using Western Blot to detect acetylation with acetyl-lysine antibody.ResultsLPS stimulated RAW264.7cells alone, the level of poly-ADP-ribosylation of HMGB1was significantly higher than the control group (0h), the level of acetylation was also significantly higher; LPS and3-AB stimulated cells, the level of poly ADP-ribosylation and acetylation are both lower than the LPS group significantly; in vitro, the poly ADP-ribosylation and acetylation level of HMGB1of the PARR-1group were significantly higher than the control group(PARR-1heat-inactivated group)(P<0.05).ConclusionPoly ADP-ribosylation of of HMGB1enhances acetylation of HMGB1。...