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  • 1
    ISSN: 1617-4623
    Keywords: Schizosaccharomyces pombe ; DNA-damage inducibility ; Damage-responsive element ; Upstream activating sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract TheSchizosaccharomyces pombe rhp51 + gene encodes a recombinational repair protein that shares significant sequence identities with the bacterial RecA and theSaccharomyces cerevisiae RAD51 protein. Levels ofrhp51 + mRNA increase following several types of DNA damage or inhibition of DNA synthesis. Anrhp51::ura4 fusion gene was used to identify the cis-acting promoter elements involved in regulatingrhp51 + expression in response to DNA damage. Two elements, designated DRE1 and DRE2 (fordamage-responsiveelement), match a decamer consensus URS (upstream repressing sequence) found in the promoters of many other DNA repair and metabolism genes fromS. cerevisiae. However, our results show that DRE1 and DRE2 each function as a UAS (upstream activating sequence) rather than a URS and are also required for DNA-damage inducibility of the gene. A 20-bp fragment located downstream of both DRE1 and DRE2 is responsible for URS function. The DRE1 and DRE2 elements cross-competed for binding to two proteins of 45 and 59 kDa. DNase I footprint analysis suggests that DRE1 and DRE2 bind to the same DNA-binding proteins. These results suggest that the DRE-binding proteins may play an important role in the DNA-damage inducibility ofrhp51 + expression.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Key words Rat ; DNA topoisomerase IIα ; Saccharomyces cerevisiae ; top2ts ; Complementation ; Leucine zipper
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  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 top2 ts . 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.
    Type of Medium: Electronic Resource
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