Negative regulation of transcription by the yeast global transcription factors, Gal11 and Sin4.
Gal11 and Sin4 proteins are yeast global transcription factors that regulate transcription of a variety of genes, both positively and negatively. Gal11, in a major part, functions in the activation of transcription, whereas Sin4 has an opposite role, yet they are reported to be present as a complex in the so-called RNA polymerase II holoenzyme. To reveal howthese auxiliary factors participate in switching transcription on and off, a complex formation between Gal11 and Sin4 and its effect on the negative regulation of transcription were studied. Using an artificial promoter that is negatively regulated by Gal11, it was shown that the presence of Sin4 or Pgd1/Hrs1/Med3 was required for Gal11 to repress both basal and activated transcription. Genetic and biochemical studies using a temperature-sensitive Gal11 mutant showed that the amino acid region 866-910 essential for Gal11 function was also important for repression of transcription and a complex formation with Sin4. Analysis with dam methylase accessibility to the promoter region suggested that nucleosome structure may be involved in negative regulation. Based on these results, possible mechanisms by which a mediator subcomplex regulates transcription is discussed.
Nishizawa
M
M
Department of Microbiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan. mas@mc.med.keio.ac.jp
D015153:Blotting, Western; D019175:DNA Methylation; D004271:DNA, Fungal / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D056892:Mediator Complex; D016296:Mutagenesis; D010957:Plasmids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000032:analysis
D004271:DNA, Fungal; D005656:Fungal Proteins; C058337:GAL11 protein, S cerevisiae; D056892:Mediator Complex; C076878:SIN4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D001616:beta-Galactosidase