Tight cooperation between Mot1p and NC2 in regulating genome-wide transcription, repression of transcription following heat shock induction and genetic interaction with SAGA

Molecular Cancer Research, Netherlands Proteomics Centre, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands.
Nucleic Acids Research (Impact Factor: 9.11). 02/2012; 40(3):996-1008. DOI: 10.1093/nar/gkr784
Source: PubMed


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.

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Article: Tight cooperation between Mot1p and NC2 in regulating genome-wide transcription, repression of transcription following heat shock induction and genetic interaction with SAGA

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    • "Cryptic promoters resemble canonical promoters in the sense that they can be SAGA- or TFIID-regulated. The FLO8 cryptic promoter contains a functional TATA box at coordinate +1626 (4), suggesting that its activity is dependent on the SAGA complex and is repressed by Mot1p and NC2 (18,30,31). Conditional depletion of Mot1p or NC2 results in an increase of intragenic levels at FLO8. "
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