Mot1p is essential for TBP recruitment to selected promoters during in vivo gene activation.
Recruitment of TATA-binding protein (TBP) is central to activation of transcription by RNA polymerase II (pol II). This depends upon co-activator proteins including TBP-associated factors (TAFs). Yeast Mot1p was identified as a general transcriptional repressor in genetic screens and is also found associated with TBP. To obtain insight into Mot1p function in vivo, we determined the mRNA expression profile of the mot1-1 temperature-sensitive (Ts) strain. Unexpectedly, this indicated that Mot1p mostly plays a positive role for transcription. For one potential activation target, HXT2, we analyzed promoter recruitment of Mot1p, TBP, Taf1p (Taf130p) and pol II by chromatin immunoprecipitation assays. Whereas TBP becomes stably associated upon activation of the HXT2 and HXT4 promoters, Mot1p showed only a transient association. TBP recruitment was compromised in two different mot1 mutant strains, but was only moderately affected in a taf1 Ts strain. Together, our data indicate that Mot1p can assist in recruitment of TBP on promoters during gene activation in vivo.
D000251:Adenosine Triphosphatases; D004265:DNA Helicases / Q000378:metabolism*; D004257:DNA Polymerase II / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D051246:Glucose Transport Proteins, Facilitative; D008565:Membrane Proteins / Q000235:genetics; D009004:Monosaccharide Transport Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D035381:TATA-Binding Protein Associated Factors / Q000378:metabolism*; D035181:TATA-Box Binding Protein / Q000378:metabolism*; D015533:Transcriptional Activation / Q000235:genetics*
D051246:Glucose Transport Proteins, Facilitative; C084498:HXT2 protein, S cerevisiae; C090593:HXT4 protein, S cerevisiae; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D004257:DNA Polymerase II; D000251:Adenosine Triphosphatases; C073676:MOT1 protein, S cerevisiae; D004265:DNA Helicases