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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...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2008
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Materias: | |
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. |
format | Text |
id | pubmed-2396426 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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