Role of UME6 in transcriptional regulation of a DNA repair gene in Saccharomyces cerevisiae.
In Saccharomyces cerevisiae UV radiation and a variety of chemical DNA-damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of these genes is PHR1, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHR1 require an upstream activation sequence, UAS(PHR1), which has homology with DRC elements found upstream of at least 19 other DNA repair and DNA metabolism genes in yeast. Here we report the identification of the UME6 gene of S. cerevisiae as a regulator of UAS(PHR1) activity. Multiple copies of UME6 stimulate expression from UAS(PHR1) and the intact PHR1 gene. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHR1 is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UME6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHR1 mRNA, and increases the UV sensitivity of a rad2 mutant. Despite the fact that UAS(PHR1) does not contain the URS1 sequence, which has been previously implicated in UME6-mediated transcriptional regulation, we find that Ume6p binds to UAS(PHR1) with an affinity and a specificity similar to those seen for a URS1 site. Similar binding is also seen for DRC elements from RAD2, RAD7, and RAD53, suggesting that UME6 contributes to the regulated expression of a subset of damage-responsive genes in yeast.
Sweet
D H
DH
Department of Biochemistry and Biophysics, School of Medicine, The University of North Carolina at Chapel Hill, 27599-7260, USA.
Jang
Y K
YK
Sancar
G B
GB
D001051:Apoenzymes / Q000096:biosynthesis / Q000235:genetics*; D001665:Binding Sites; D018797:Cell Cycle Proteins; D064447:Checkpoint Kinase 2; D003001:Cloning, Molecular; D004260:DNA Repair / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000096:biosynthesis / Q000235:genetics*; D004706:Endodeoxyribonucleases; D005656:Fungal Proteins / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D008562:Membrane Glycoproteins; D016296:Mutagenesis; D011485:Protein Binding; D011494:Protein Kinases / Q000235:genetics; D017346:Protein Serine-Threonine Kinases; D011740:Pyrimidine Dimers / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D012097:Repressor Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014466:Ultraviolet Rays / Q000009:adverse effects
D001051:Apoenzymes; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; C091515:PHR1 protein, Candida albicans; D011740:Pyrimidine Dimers; C063915:RAD7 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C089979:UME6 protein, S cerevisiae; C073998:RAD2 protein, S cerevisiae; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; C067022:RAD53 protein, S cerevisiae; D004706:Endodeoxyribonucleases; D004255:Deoxyribodipyrimidine Photo-Lyase