Activation of ADP-ribosyltransferase in polyamine-depleted mammalian cells.
Mammalian fibroblasts were cultured in the presence of alpha-methylornithine and/or methylglyoxal bis(guanylhydrazone), which inhibit the synthesis of polyamines. This led to a decrease in the cellular content of the polyamines spermine and spermidine by up to 60% when the cells were grown in the presence of both drugs together. The activity of the chromatin-associated enzyme ADP-ribosyltransferase was enhanced 2-3-fold in the drug-treated cells when measured in cells subsequently rendered permeable to exogenous NAD+, the substrate for the transferase. This is a novel and surprising observation, since the transferase is invariably activated by the addition of polyamines to a suitable incubation system such as permeabilized cells, isolated nuclei or the purified enzyme. We found no evidence that the activation was due to the appearance of DNA strand breaks, by using a variety of procedures including both neutral [the 'nucleoid' technique of Cook & Brazell [(1975) J. Cell Sci. 19, 261-279; (1976) J. Cell Sci. 22, 287-302]] and alkaline sucrose-gradient centrifugation and gel electrophoresis, suggesting that this therefore may not be the only means of regulating the activity of ADP-ribosyltransferase and that polyamines may have a role to play in this regard in vivo.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D007668:Kidney; D008935:Mitoguazone / Q000494:pharmacology; D009713:Nucleotidyltransferases / Q000378:metabolism*; D009952:Ornithine / Q000031:analogs & derivatives / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases; D013095:Spermidine / Q000378:metabolism*; D013096:Spermine / Q000378:metabolism*
D000246:Adenosine Diphosphate Ribose; D013096:Spermine; C025025:alpha-methylornithine; D004247:DNA; D009952:Ornithine; D011065:Poly(ADP-ribose) Polymerases; D009713:Nucleotidyltransferases; D008935:Mitoguazone; D013095:Spermidine