Expression of human poly(ADP-ribose) polymerase in Saccharomyces cerevisiae.
The coding sequence for human poly(ADP-ribose) polymerase was expressed inducibly in Saccharomyces cerevisiae from a low-copy-number plasmid vector. Cell free extracts of induced cells had poly(ADP-ribose) polymerase activity when assayed under standard conditions; activity could not be detected in noninduced cell extracts. Induced cells formed poly(ADP-ribose) in vivo, and levels of these polymers increased when cells were treated with the alkylating agent N-methyl-N'-nitro-N- nitrosoguanidine (MNNG). The cytotoxicity of this agent was increased in induced cells, and in vivo labelling with [3H]adenine further decreased their viability. Increased levels of poly(ADP-ribose) found in cells treated with the alkylating agent were not accompanied by lowering of the NAD concentration.
Collinge
M A
MA
Institute of Pharmacology and Biochemistry, University of Zurich, Tierspital, Switzerland.
Althaus
F R
FR
D002470:Cell Survival / Q000187:drug effects; D002474:Cell-Free System; D015870:Gene Expression; D006801:Humans; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D008769:Methylnitronitrosoguanidine; D011065:Poly(ADP-ribose) Polymerases