title: Specific defects in different transcription complexes compensate for the requirement of the negative cofactor 2 repressor in Saccharomyces cerevisiae.
abstract: Negative cofactor 2 (NC2) has been described as an essential and evolutionarily conserved transcriptional repressor, although in vitro and in vivo experiments suggest that it can function as both a positive and a negative effector of transcription. NC2 operates by interacting with the core promoter and components of the basal transcription machinery, like the TATA-binding protein (TBP). In this work, we have isolated mutants that suppress the growth defect caused by the depletion of NC2. We have identified mutations affecting components of three different complexes involved in the control of basal transcription: the mediator, TFIIH, and RNA pol II itself. Mutations in RNA pol II include both overexpression of truncated forms of the two largest subunits (Rpb1 and Rpb2) and reduced levels of these proteins. Suppression of NC2 depletion was also observed by reducing the amounts of the mediator essential components Nut2 and Med7, as well as by deleting any of the nonessential mediator components, except Med2, Med3, and Gal11 subunits. Interestingly, the Med2/Med3/Gal11 triad forms a submodule within the mediator tail. Our results support the existence of different components within the basic transcription complexes that antagonistically interact with the NC2 repressor and suggest that the correct balance between the activities of specific positive and negative components is essential for cell growth.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Valencia, 46100 Burjassot, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Peiró-Chova
authors.forename: Francisco
authors.initials: F
authors.lastname: Estruch
mesh_terms: D000251:Adenosine Triphosphatases; D000483:Alleles; D004265:DNA Helicases / Q000172:deficiency; D050505:Mutant Proteins / Q000302:isolation & purification; D009154:Mutation / Q000235:genetics; D010750:Phosphoproteins / Q000172:deficiency / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D021122:Protein Subunits / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D035381:TATA-Binding Protein Associated Factors / Q000172:deficiency; D015534:Trans-Activators / Q000235:genetics; D035581:Transcription Factor TFIIB / Q000378:metabolism; D035362:Transcription Factor TFIID / Q000378:metabolism; D051758:Transcription Factor TFIIH / Q000378:metabolism; D014157:Transcription Factors / Q000172:deficiency / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: C518355:GAL10 protein, S cerevisiae; D050505:Mutant Proteins; D010750:Phosphoproteins; D021122:Protein Subunits; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D035381:TATA-Binding Protein Associated Factors; D015534:Trans-Activators; D035581:Transcription Factor TFIIB; D035362:Transcription Factor TFIID; D014157:Transcription Factors; C075995:down-regulator of transcription 1; D051758:Transcription Factor TFIIH; D012319:RNA Polymerase II; D000251:Adenosine Triphosphatases; C073676:MOT1 protein, S cerevisiae; D004265:DNA Helicases