A novel cis-acting cysteine-responsive regulatory element of the gene for the high-affinity glutathione transporter of Saccharomyces cerevisiae.
We cloned a DNA fragment from Saccharomyces cerevisiae that complemented the deficiency in high-affinity glutathione transport activity conferred by a gsh11 mutation, and found that the ORF responsible was YJL212c, which had already been designated as OPT1 and HGT1 by others. Northern analysis clearly demonstrated that this ORF, now referred to as OPT1/ HGT1/ GSH11, was induced by sulfur starvation and repressed by adding cysteine to the growth medium. Reporter gene assays showed that a segment spanning the region between positions -371 and -355 was essential for the regulation of this gene. A sequence of 9 nt, CCGCCACAC (from -364 to -356), in this region was shown to be required for protein binding, using an electrophoretic mobility shift assay. Based on these results, we propose that CCGCCACAC comprises the core of a cis-acting element involved in cysteine-responsive gene regulation in S. cerevisiae.
Miyake
T
T
Industrial Technology Center of Okayama Prefecture, 5301 Haga, Okayama 701-1296, Japan. tsuyoshi_miyake@pref.okayama.jp
Kanayama
M
M
Sammoto
H
H
Ono
B
B
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism / Q000494:pharmacology*; D004271:DNA, Fungal / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology; D026901:Membrane Transport Proteins; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development
D002352:Carrier Proteins; D004271:DNA, Fungal; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; C086860:glutathione transporter; D005978:Glutathione; D003545:Cysteine