Selective isolation of domains of chromatin proximal to both carcinogen-induced DNA damage and poly-adenosine diphosphate-ribosylation.
Poly-adenosine diphosphate (ADP)-ribosylation of nuclear proteins has been demonstrated previously to be activated in vivo by the presence of DNA single-strand breaks and has thus been implicated to play an important role in altering chromatin structure during cellular recovery from DNA damage. Based upon these considerations, a novel immunofractionation method, using antipoly(ADP-ribose) coupled to Sepharose, has been used to enrich for those limited domains of chromatin undergoing poly-ADP-ribosylation. We have used three independent methods to verify the presence of significant levels of single-strand DNA breaks adjacent to polynucleosomes engaged in ADP-ribosylation.
D002843:Chromatin / Q000302:isolation & purification*; D003561:Cytarabine / Q000378:metabolism; D004247:DNA / Q000032:analysis*; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects; D006367:HeLa Cells / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008770:Methylnitrosourea / Q000633:toxicity*; D009243:NAD / Q000378:metabolism; D009607:Nitrosourea Compounds / Q000633:toxicity*; D009702:Nucleoside Diphosphate Sugars / Q000032:analysis*; D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D014316:Tritium
D002843:Chromatin; D009607:Nitrosourea Compounds; D009702:Nucleoside Diphosphate Sugars; D010761:Phosphorus Radioisotopes; D003561:Cytarabine; D009243:NAD; D014316:Tritium; D011064:Poly Adenosine Diphosphate Ribose; D008770:Methylnitrosourea; D004247:DNA; D011065:Poly(ADP-ribose) Polymerases