An artificial transcription activator mimics the genome-wide properties of the yeast Pdr1 transcription factor.
We analysed the genome-wide regulatory properties of an artificial transcription activator in which the DNA-binding domain of the yeast transcription factor, Pdr1, was fused to the activation domain of Gal4 (Pdr1*GAD). This Pdr1*GAD chimera was put under the control of the inducible GAL1 promoter. DNA microarray analyses showed that all the target genes upregulated by the well-studied native gain-of-function Pdr1-3 mutant were similarly activated by the chimerical factor Pdr1*GAD upon galactose induction. Additionally, this kinetic approach led us not only to confirm previously published targets, but also to define a hierarchy among members of the Pdr1 regulon. Our observations prove, for the first time at the complete genome level, that the DNA-binding domain of Pdr1 is sufficient to guide its specificity. We propose that this approach could be useful for the study of new transcription factors identified in silico from sequenced organisms. Complete data are available at www.biologie.ens.fr/yeast-publi.html.
Devaux
F
F
Laboratoire de Génétique Moléculaire, CNRS UMR 8541, Ecole Normale Supérieure, 46 rue d'Ulm, Paris, France.
Marc
P
P
Bouchoux
C
C
Delaveau
T
T
Hikkel
I
I
Potier
M C
MC
Jacq
C
C
D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics*; D005690:Galactose / Q000378:metabolism; D016678:Genome; D007700:Kinetics; D008957:Models, Genetic; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors / Q000737:chemistry; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D015854:Up-Regulation
D004268:DNA-Binding Proteins; C085480:PDR1 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D005690:Galactose