The activity of Adenosine diphosphoribosyl transferase (ADPRT), the chromatin-bound enzyme which specifically catalyses the cleavage of oxidized NAD<sup>+</sup> 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)<sub>n</sub> 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)<sub>n</sub> chain length by both cell types. Polyamines which are present at high concentrations in rapidly dividing tissues were able to stimulate ADPRT activity both <i>in vitro</i> and <i>in vivo</i> 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)<sub>n</sub> 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 <i>in vivo</i>.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:377600 |
| Date | January 1986 |
| Creators | Gordon, A. M. |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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