title: Tight cooperation between Mot1p and NC2β in regulating genome-wide transcription, repression of transcription following heat shock induction and genetic interaction with SAGA.
abstract: TATA-binding protein (TBP) is central to the regulation of eukaryotic transcription initiation. Recruitment of TBP to target genes can be positively regulated by one of two basal transcription factor complexes: SAGA or TFIID. Negative regulation of TBP promoter association can be performed by Mot1p or the NC2 complex. Recent evidence suggests that Mot1p, NC2 and TBP form a DNA-dependent protein complex. Here, we compare the functions of Mot1p and NC2βduring basal and activated transcription using the anchor-away technique for conditional nuclear depletion. Genome-wide expression analysis indicates that both proteins regulate a highly similar set of genes. Upregulated genes were enriched for SAGA occupancy, while downregulated genes preferred TFIID binding. Mot1p and NC2β depletion during heat shock resulted in failure to downregulate gene expression after initial activation, which was accompanied by increased TBP and RNA pol II promoter occupancies. Depletion of Mot1p or NC2β displayed preferential synthetic lethality with the TBP-interaction module of SAGA. Our results support the model that Mot1p and NC2β directly cooperate in vivo to regulate TBP function, and that they are involved in maintaining basal expression levels as well as in resetting gene expression after induction by stress.
authors: Molecular Cancer Research, Netherlands Proteomics Centre, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands.
authors: Gianpiero
authors: G
authors: Spedale
authors: Claartje A
authors: CA
authors: Meddens
authors: Maria J E
authors: MJ
authors: Koster
authors: Cheuk W
authors: CW
authors: Ko
authors: Sander R
authors: SR
authors: van Hooff
authors: Frank C P
authors: FC
authors: Holstege
authors: H Th Marc
authors: HT
authors: Timmers
authors: W W M Pim
authors: WW
authors: Pijnappel
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D006360:Heat-Shock Proteins / Q000378:metabolism; D018869:Heat-Shock Response; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D035381:TATA-Binding Protein Associated Factors / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: C499528:HSP26 protein, S cerevisiae; D006360:Heat-Shock Proteins; D012097:Repressor Proteins; C507785:SAGA complex, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D035381:TATA-Binding Protein Associated Factors; D015534:Trans-Activators; D012319:RNA Polymerase II; D000251:Adenosine Triphosphatases; C073676:MOT1 protein, S cerevisiae