Requirement of multiple DNA-protein interactions for inducible expression of RNR3 gene in Saccharomyces cerevisiae in response to DNA damage.
The RNR3 gene encodes the large subunit of ribonucleotide reductase. Transcription of this gene is induced 12-fold in response to DNA damage or by a DNA replication blocker. To investigate cis-acting regulation, deletion analysis of the promoter region of the RNR3 gene was performed and we identified two upstream-repressing sequences in the RNR3 regulatory region. An 18-base-pairs fragment, termed DNA-damage responsive element 1 (DRE1) located between -212 and -194 in this region was found to be essential for the induction of RNR3. This fragment contained a negatively acting sequence where a protein factor bound to the region during normal growth but disappeared by exposure to 4-nitroquinoline-1-oxide. The other repressive element homologue to DRE1 was located at -263 to -254. One possible upstream-activating sequence which regulates the basal expression of RNR3 was also found. These results show that at least three potential cis-elements are necessary for the inducible expression of yeast expression of yeast RNR3 in response to DNA damage.
Endo-Ichikawa
Y
Y
Department of Public Health, Kansai Medical University, Osaka, Japan.
Kohno
H
H
Furukawa
T
T
Ueda
T
T
Ogawa
Y
Y
Tokunaga
R
R
Taketani
S
S
D015112:4-Nitroquinoline-1-oxide / Q000494:pharmacology; D001482:Base Composition; D001483:Base Sequence; D004249:DNA Damage; D017931:DNA Primers; D015342:DNA Probes; D004261:DNA Replication; D004271:DNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D004790:Enzyme Induction; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal; D006918:Hydroxyurea / Q000494:pharmacology; D046911:Macromolecular Substances; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D009153:Mutagens / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D012264:Ribonucleotide Reductases / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D017384:Sequence Deletion; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis
D017931:DNA Primers; D015342:DNA Probes; D004271:DNA, Fungal; D046911:Macromolecular Substances; D009153:Mutagens; D011993:Recombinant Fusion Proteins; D008769:Methylnitronitrosoguanidine; D015112:4-Nitroquinoline-1-oxide; D012264:Ribonucleotide Reductases; D001616:beta-Galactosidase; D006918:Hydroxyurea