Yap1 overproduction restores arsenite resistance to the ABC transporter deficient mutant ycf1 by activating ACR3 expression.
Ycf1 and Acr3 are transporters that have been previously shown to protect Saccharomyces cerevisiae cells from the toxic effects of arsenite. Ycf1 and Acr3 are positively regulated by distinct, but related bZIP transcriptional activators, Yap1 and Yap8, respectively. In this study, we show that overexpression of Yap1 complemented the arsenite hypersensitivity of the ycf1 null mutant, but only if the ACR3 gene is functional. We further show that the expression of either an ACR3-lacZ promoter fusion reporter or the endogenous ACR3 gene was stimulated by the overproduction of Yap1 upon exposure to arsenite. These data suggest that Yap1 confers arsenite resistance to the ycf1 null mutant by activating expression of the Yap8-dependent target gene, ACR3. Our data also show Yap8-dependent ACR3-lacZ expression was greatly stimulated by arsenite in a dose-dependent manner in the parental strain. However, overproduction of Yap1 in the parental strain severely limited dose-dependent activation of the reporter by arsenite. We conclude that Yap1 may compete with Yap8 for binding to the ACR3 promoter, but is unable to act as a potent activator.
Bouganim
N
N
University of Montreal, Guy-Bernier Research Centre, QC, Canada.
David
J
J
Wysocki
R
R
Ramotar
D
D
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000502:physiology*; D000595:Amino Acid Sequence; D018053:Arsenites / Q000494:pharmacology*; D015152:Blotting, Northern; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics*; D005786:Gene Expression Regulation / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000096:biosynthesis* / Q000737:chemistry / Q000502:physiology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D018053:Arsenites; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C086677:YAP1 protein, S cerevisiae; C089179:YCF1 protein, S cerevisiae; D001616:beta-Galactosidase; C015001:arsenite