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TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages

Transcription in eukaryotes is a multistep process involving the assembly and disassembly of numerous inter- and intramolecular interactions between transcription factors and nucleic acids. The roles of each of these interactions and the regions responsible for them have been identified and studied...

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Detalles Bibliográficos
Autores principales: Sevilimedu, Aarti, Shi, Hua, Lis, John T.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2396426/
https://www.ncbi.nlm.nih.gov/pubmed/18403417
http://dx.doi.org/10.1093/nar/gkn163
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author Sevilimedu, Aarti
Shi, Hua
Lis, John T.
author_facet Sevilimedu, Aarti
Shi, Hua
Lis, John T.
author_sort Sevilimedu, Aarti
collection PubMed
description Transcription in eukaryotes is a multistep process involving the assembly and disassembly of numerous inter- and intramolecular interactions between transcription factors and nucleic acids. The roles of each of these interactions and the regions responsible for them have been identified and studied primarily by the use of mutants, which destroy the inherent properties of the interacting surface. A less intrusive but potentially effective way to study the interactions as well as the surfaces responsible for them is the use of RNA aptamers that bind to the interacting factors. Here, we report the isolation and characterization of high-affinity RNA aptamers that bind to the yeast general transcription factor TFIIB. These aptamers fall into two classes that interfere with TFIIB's interactions with either TBP or RNA polymerase II, both of which are crucial for transcription in yeast. We demonstrate the high affinity and specificity of these reagents, their effect on transcription and preinitiation complex formation and discuss their potential use to address mechanistic questions in vitro as well as in vivo.
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spelling pubmed-23964262008-05-28 TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages Sevilimedu, Aarti Shi, Hua Lis, John T. Nucleic Acids Res RNA Transcription in eukaryotes is a multistep process involving the assembly and disassembly of numerous inter- and intramolecular interactions between transcription factors and nucleic acids. The roles of each of these interactions and the regions responsible for them have been identified and studied primarily by the use of mutants, which destroy the inherent properties of the interacting surface. A less intrusive but potentially effective way to study the interactions as well as the surfaces responsible for them is the use of RNA aptamers that bind to the interacting factors. Here, we report the isolation and characterization of high-affinity RNA aptamers that bind to the yeast general transcription factor TFIIB. These aptamers fall into two classes that interfere with TFIIB's interactions with either TBP or RNA polymerase II, both of which are crucial for transcription in yeast. We demonstrate the high affinity and specificity of these reagents, their effect on transcription and preinitiation complex formation and discuss their potential use to address mechanistic questions in vitro as well as in vivo. Oxford University Press 2008-05 2008-04-10 /pmc/articles/PMC2396426/ /pubmed/18403417 http://dx.doi.org/10.1093/nar/gkn163 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 RNA
Sevilimedu, Aarti
Shi, Hua
Lis, John T.
TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages
title TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages
title_full TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages
title_fullStr TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages
title_full_unstemmed TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages
title_short TFIIB aptamers inhibit transcription by perturbing PIC formation at distinct stages
title_sort tfiib aptamers inhibit transcription by perturbing pic formation at distinct stages
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2396426/
https://www.ncbi.nlm.nih.gov/pubmed/18403417
http://dx.doi.org/10.1093/nar/gkn163
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