DNA repair-associated ADP-ribosylation in vivo. Modification of histone H1 differs from that of the principal acceptor proteins.
ADP-ribosylation in vivo of histone H1 was studied in hepatoma cells (Yoshida AH 7974) after treatment with the alkylating agent dimethyl sulfate for 30 min and compared with that of other polypeptides. In unstimulated cells, histone H1 was only a minor acceptor (less than 4%) of total monomeric and polymeric ADP-ribosyl residues. Induction of DNA repair by dimethyl sulfate treatment increased total mono(ADP-ribosyl) protein conjugates 1.6-fold whereas histone H1-linked mono(ADP-ribosyl) groups were elevated greater than 30-fold, thus accounting for nearly one-fourth of the net increase in monomeric ADP-ribosyl residues. In contrast, histone H1-associated poly(ADP-ribosyl) residues comprised only 2% of the total increase in poly(ADP-ribose). The extent to which the histone H1 population became ADP-ribosylated was low even in dimethyl sulfate-treated cells. Less than 2% of the histone H1 molecules were mono(ADP-ribosyl)ated and only 0.003% carried poly(ADP-ribosyl) chains when an average chain length of 10 is assumed. The principal polypeptide acceptors of alkylation-induced ADP-ribosylation were concentrated in two peaks, one migrating close to the position of core histones H3/H2B and accepting most of the induced mono(ADP-ribosyl) and poly(ADP-ribosyl) residues. The other (Mr = 110,000-160,000) resembled auto-modified poly(ADP-ribose) polymerase. Our data demonstrate marked differences of alkylation-induced (ADP-ribosyl)n protein patterns to analyses performed in vitro.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000478:Alkylation; D000818:Animals; D004260:DNA Repair; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006657:Histones / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D008970:Molecular Weight; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013463:Sulfuric Acid Esters / Q000494:pharmacology
D006657:Histones; D009702:Nucleoside Diphosphate Sugars; D013463:Sulfuric Acid Esters; D000246:Adenosine Diphosphate Ribose; D011064:Poly Adenosine Diphosphate Ribose; C007482:dimethyl sulfate