A study of poly(ADP-ribosyl)ation in polyoma virus-transformed and untransformed BHK21/C13 cells
The activity of Adenosine diphosphoribosyl transferase (ADPRT), the chromatin-bound enzyme which specifically catalyses the cleavage of oxidized NAD+ with the concomitant covalent attachment of the ADP-ribose moiety to acceptor proteins, was investigated in Polyoma Virus-Transformed (PyY) and Untransformed BHK21/C13 (BHK) cells. It was shown that ADPRT activity was consistently 2-4 fold higher in PyY cells than in BHK cells. The spectrum of (ADP-ribose)n residues synthesised by the two cell types was very similar when analysed by hydroxyapatite column chromatography. Poly (ADP-ribose) Glycohydrolase activity in the two cell types was identical with 25-30% degradation of the poly(ADP-ribose) over a period of 90 minutes. DNA damage resulting from incubation with Deoxyribonuclease was reflected by an immediate increase in ADPRT activity and an increase in (ADP-ribose)n chain length by both cell types. Polyamines which are present at high concentrations in rapidly dividing tissues were able to stimulate ADPRT activity both in vitro and in vivo in BHK and PyY cells. In general the average chain length of ADP-ribose residues synthesised remained unaltered. No significant increase in the level of DNA-strand breakage could be detected in the polyamine-treated cells. Depletion of the cellular polyamine levels resulted in stimulation of ADPRT activity, but there was no significant difference in the spectrum of (ADP-ribose)n residues synthesised. Again no significant increase in the level of DNA-strand breaks could be detected in the polyamine-depleted cells. These results suggest that DNA-damage may not be the only means of regulating ADPRT activity and that polyamines may have a role to play in this regard in vivo.