The effect of in vivo treatment with triiodothyronine on the in vitro synthesis of protein-poly(ADP)-ribose adducts by isolated cardiocyte nuclei and the separation of poly(ADP)-ribosylated proteins by phenol extraction and electrophoresis.

Covalent binding of 4-carbamoylbenzenediazonium chloride to deoxyguanine bases of DNA resulting in apparent irreversible inhibition of poly(adenosine diphosphoribose) polymerase at the nicotinamide binding site

“…No 3H was detected in gel slices, indicating only ADP-ribose labeling. (C) A suspension (10 mL) of liver nuclei (2 mg/mL protein) was allowed to react with [3,5-3H2]-4-diazoniobenzamide (5.1 pd, 0.1 mM) at 4 °C for 30 min, and the reaction was terminated as described for (A), followed by gel electrophoresis (Jackowski & Kun, 1983). (D) Electrophoretogram illustrating the decay of DNA-bound [3,5-3H2]-4-diazoniobenzamide following digestion with 10 Mg of DNase I for 30 min at 25 °C.…”

“…Gel Electrophoresis. Gel electrophoresis was performed according to the procedure of Jackowski and Kun (1983).…”

“…However, the infrequent association of the enzyme protein with DNA termini (DeMurcia et al, 1983;Ittel et al, 1985) seems inconsistent with a hypothesis that correlates DNA fragmentation with increased coenzymic activity. Furthermore, a high degree of DNA fragmentation was shown to coincide with low enzymatic activity in certain cell types (Skidmore et al, 1985); thus activation of poly-ADPribosylations by DNA fragmentation is not universal (Jackowski & Kun, 1983). It was concluded on the basis of enzymatic experiments with synthetic oligodeoxyribonucleotides (Berger & Petzold, 1985) that small coenzymatically active DNA fragments may exist in nuclei which cannot be detected by tests suitable for the determination of DNA fragmentation.…”