title: Acetyl coenzyme A stimulates RNA polymerase II transcription and promoter binding by transcription factor IID in the absence of histones.
abstract: Protein acetylation has emerged as a means of controlling levels of mRNA synthesis in eukaryotic cells. Here we report that acetyl coenzyme A (acetyl-CoA) stimulates RNA polymerase II transcription in vitro in the absence of histones. The effect of acetyl-CoA on basal and activated transcription was studied in a human RNA polymerase II transcription system reconstituted from recombinant and highly purified transcription factors. Both basal and activated transcription were stimulated by the addition of acetyl-CoA to transcription reaction mixtures. By varying the concentrations of general transcription factors in the reaction mixtures, we found that acetyl-CoA decreased the concentration of TFIID required to observe transcription. Electrophoretic mobility shift assays and DNase I footprinting revealed that acetyl-CoA increased the affinity of the general transcription factor TFIID for promoter DNA in a TBP-associated factor (TAF)-dependent manner. Interestingly, acetyl-CoA also caused a conformational change in the TFIID-TFIIA-promoter complex as assessed by DNase I footprinting. These results show that acetyl-CoA alters the DNA binding activity of TFIID and indicate that this biologically important cofactor functions at multiple levels to control gene expression.
authors.affiliation: Department of Chemistry, University of Colorado at Boulder, Boulder, Colorado 80309-0215, USA.
authors.forename: S K
authors.initials: SK
authors.lastname: Galasinski
authors.forename: T N
authors.initials: TN
authors.lastname: Lively
authors.forename: A
authors.initials: A
authors.lastname: Grebe De Barron
authors.forename: J A
authors.initials: JA
authors.lastname: Goodrich
mesh_terms: D000105:Acetyl Coenzyme A / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D035362:Transcription Factor TFIID; D019473:Transcription Factors, TFII / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D006657:Histones; D035362:Transcription Factor TFIID; D019473:Transcription Factors, TFII; D000105:Acetyl Coenzyme A; D012319:RNA Polymerase II