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Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box

In Saccharomyces cerevisiae, multiple approaches have arrived at a consensus TATA box sequence of TATA(T/A)A(A/T)(A/G). TATA-binding protein (TBP) affinity alone does not determine TATA box function. To discover how a minimal set of factors required for basal and activated transcription contributed...

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Autores principales: Bjornsdottir, Gudrun, Myers, Lawrence C.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2396422/
https://www.ncbi.nlm.nih.gov/pubmed/18385157
http://dx.doi.org/10.1093/nar/gkn130
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author Bjornsdottir, Gudrun
Myers, Lawrence C.
author_facet Bjornsdottir, Gudrun
Myers, Lawrence C.
author_sort Bjornsdottir, Gudrun
collection PubMed
description In Saccharomyces cerevisiae, multiple approaches have arrived at a consensus TATA box sequence of TATA(T/A)A(A/T)(A/G). TATA-binding protein (TBP) affinity alone does not determine TATA box function. To discover how a minimal set of factors required for basal and activated transcription contributed to the sequence requirements for a functional TATA box, we performed transcription reactions using highly purified proteins and CYC1 promoter TATA box mutants. The TATA box consensus sequence is a good predictor of promoter activity. However, several nonconsensus sequences are almost fully functional, indicating that mechanistic requirements are not the only selective pressure on the TATA box. We also found that the effect of a mutation at a certain position is often dependent on other bases within a particular TATA box. Although activators and coactivators strongly influence TBP recruitment and stability at promoters, neither Mediator, the activator Gal4-V16, nor TFIID specifically compensate for the low transcription levels of the weak TATA boxes. The addition of Mediator to purified transcription reactions did, however, increase the functional selectivity for certain consensus TATA sequences. Transcription in whole-cell extracts or in vivo with these TATA box mutants indicated that factors, other than those in our purified system, may help initiate transcription from weak TATA boxes.
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spelling pubmed-23964222008-05-28 Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box Bjornsdottir, Gudrun Myers, Lawrence C. Nucleic Acids Res Molecular Biology In Saccharomyces cerevisiae, multiple approaches have arrived at a consensus TATA box sequence of TATA(T/A)A(A/T)(A/G). TATA-binding protein (TBP) affinity alone does not determine TATA box function. To discover how a minimal set of factors required for basal and activated transcription contributed to the sequence requirements for a functional TATA box, we performed transcription reactions using highly purified proteins and CYC1 promoter TATA box mutants. The TATA box consensus sequence is a good predictor of promoter activity. However, several nonconsensus sequences are almost fully functional, indicating that mechanistic requirements are not the only selective pressure on the TATA box. We also found that the effect of a mutation at a certain position is often dependent on other bases within a particular TATA box. Although activators and coactivators strongly influence TBP recruitment and stability at promoters, neither Mediator, the activator Gal4-V16, nor TFIID specifically compensate for the low transcription levels of the weak TATA boxes. The addition of Mediator to purified transcription reactions did, however, increase the functional selectivity for certain consensus TATA sequences. Transcription in whole-cell extracts or in vivo with these TATA box mutants indicated that factors, other than those in our purified system, may help initiate transcription from weak TATA boxes. Oxford University Press 2008-05 2008-04-01 /pmc/articles/PMC2396422/ /pubmed/18385157 http://dx.doi.org/10.1093/nar/gkn130 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Molecular Biology
Bjornsdottir, Gudrun
Myers, Lawrence C.
Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box
title Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box
title_full Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box
title_fullStr Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box
title_full_unstemmed Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box
title_short Minimal components of the RNA polymerase II transcription apparatus determine the consensus TATA box
title_sort minimal components of the rna polymerase ii transcription apparatus determine the consensus tata box
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2396422/
https://www.ncbi.nlm.nih.gov/pubmed/18385157
http://dx.doi.org/10.1093/nar/gkn130
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