Expression of human poly(ADP-ribose) polymerase in Saccharomyces cerevisiae |
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Authors: | M. A. Collinge F. R. Althaus |
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Affiliation: | 1. Institute of Pharmacology and Biochemistry, University of Zurich, Tierspital, Winterthurerstrasse 260, 8057, Zürich, Switzerland
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Abstract: | 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(ADPribose) polymerase activity when assayed under standard conditions; activity could not be detected in non-induced 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. |
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