A series of yeast expression plasmids which comprise segments of the cDNA sequences encoding rat topo IIα have been constructed. The transcription of these constructs is under the control of the yeast GAL1 promoter. Galactose-dependent expression of the cloned rat topo IIα cDNA complemented a yeast top2 ts mutation, as well as a deletion mutation at the yeast TOP2 locus. Truncation of 12 N-terminal amino acids and/or 158 C-terminal amino acids of rat topo IIα had no effect on its ability functionally to substitute for top2ts. Moreover, a cDNA construct with mutated putative leucine zipper domain (amino acids 993–1013) retained the complementation activity. These observations suggest that transformants capable of conditional topo IIα expression can be exploited as a useful model system for studies on the structure-function relationships of wild-type and mutated topo IIα, as well as the interplay of potential antitumor drugs with the enzyme.
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Received: 4 April 1996/Accepted: 15 July 1996
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Yoon, J., Park, S., Cho, H. et al. Complementation of a yeast top2 ts mutation by a cDNA encoding rat DNA topoisomerase IIα. Mol Gen Genet 253, 81–88 (1996). https://doi.org/10.1007/s004380050299
- Key words Rat
- DNA topoisomerase IIα
- Saccharomyces cerevisiae
- Leucine zipper