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Molecular and Cellular Biology, March 2000, p. 1923-1930, Vol. 20, No. 6
Department of Chemistry and Biochemistry,
University of Colorado at Boulder, Boulder, Colorado
80309-0215,1 and Molecular Genetics
Unit, San Raffaele Biomedical Science Park, 20132 Milan,
Italy2
Received 23 November 1999/Accepted 15 December 1999
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.
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Acetyl Coenzyme A Stimulates RNA Polymerase II Transcription
and Promoter Binding by Transcription Factor IID in the
Absence of Histones
*
Corresponding author. Mailing address: Department of
Chemistry and Biochemistry, University of Colorado at Boulder, Campus Box 215, Boulder, CO 80309-0215. Phone: (303) 492-3273. Fax: (303) 492-5894. E-mail: james.goodrich{at}colorado.edu.
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