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Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25
The N-terminal acidic transactivation domain (TAD) of ERM/ETV5 (ERM(38–68)), a PEA3 group member of Ets-related transcription factors, directly interacts with the ACID/PTOV domain of the Mediator complex subunit MED25. Molecular details of this interaction were investigated using nuclear magnetic re...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4538835/ https://www.ncbi.nlm.nih.gov/pubmed/26130716 http://dx.doi.org/10.1093/nar/gkv650 |
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author | Landrieu, Isabelle Verger, Alexis Baert, Jean-Luc Rucktooa, Prakash Cantrelle, François-Xavier Dewitte, Frédérique Ferreira, Elisabeth Lens, Zoé Villeret, Vincent Monté, Didier |
author_facet | Landrieu, Isabelle Verger, Alexis Baert, Jean-Luc Rucktooa, Prakash Cantrelle, François-Xavier Dewitte, Frédérique Ferreira, Elisabeth Lens, Zoé Villeret, Vincent Monté, Didier |
author_sort | Landrieu, Isabelle |
collection | PubMed |
description | The N-terminal acidic transactivation domain (TAD) of ERM/ETV5 (ERM(38–68)), a PEA3 group member of Ets-related transcription factors, directly interacts with the ACID/PTOV domain of the Mediator complex subunit MED25. Molecular details of this interaction were investigated using nuclear magnetic resonance (NMR) spectroscopy. The TAD is disordered in solution but has a propensity to adopt local transient secondary structure. We show that it folds upon binding to MED25 and that the resulting ERM–MED25 complex displays characteristics of a fuzzy complex. Mutational analysis further reveals that two aromatic residues in the ERM TAD (F47 and W57) are involved in the binding to MED25 and participate in the ability of ERM TAD to activate transcription. Mutation of a key residue Q451 in the VP16 H1 binding pocket of MED25 affects the binding of ERM. Furthermore, competition experiments show that ERM and VP16 H1 share a common binding interface on MED25. NMR data confirms the occupancy of this binding pocket by ERM TAD. Based on these experimental data, a structural model of a functional interaction is proposed. This study provides mechanistic insights into the Mediator–transactivator interactions. |
format | Online Article Text |
id | pubmed-4538835 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-45388352015-08-18 Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 Landrieu, Isabelle Verger, Alexis Baert, Jean-Luc Rucktooa, Prakash Cantrelle, François-Xavier Dewitte, Frédérique Ferreira, Elisabeth Lens, Zoé Villeret, Vincent Monté, Didier Nucleic Acids Res Structural Biology The N-terminal acidic transactivation domain (TAD) of ERM/ETV5 (ERM(38–68)), a PEA3 group member of Ets-related transcription factors, directly interacts with the ACID/PTOV domain of the Mediator complex subunit MED25. Molecular details of this interaction were investigated using nuclear magnetic resonance (NMR) spectroscopy. The TAD is disordered in solution but has a propensity to adopt local transient secondary structure. We show that it folds upon binding to MED25 and that the resulting ERM–MED25 complex displays characteristics of a fuzzy complex. Mutational analysis further reveals that two aromatic residues in the ERM TAD (F47 and W57) are involved in the binding to MED25 and participate in the ability of ERM TAD to activate transcription. Mutation of a key residue Q451 in the VP16 H1 binding pocket of MED25 affects the binding of ERM. Furthermore, competition experiments show that ERM and VP16 H1 share a common binding interface on MED25. NMR data confirms the occupancy of this binding pocket by ERM TAD. Based on these experimental data, a structural model of a functional interaction is proposed. This study provides mechanistic insights into the Mediator–transactivator interactions. Oxford University Press 2015-08-18 2015-06-29 /pmc/articles/PMC4538835/ /pubmed/26130716 http://dx.doi.org/10.1093/nar/gkv650 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Structural Biology Landrieu, Isabelle Verger, Alexis Baert, Jean-Luc Rucktooa, Prakash Cantrelle, François-Xavier Dewitte, Frédérique Ferreira, Elisabeth Lens, Zoé Villeret, Vincent Monté, Didier Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 |
title | Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 |
title_full | Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 |
title_fullStr | Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 |
title_full_unstemmed | Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 |
title_short | Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25 |
title_sort | characterization of erm transactivation domain binding to the acid/ptov domain of the mediator subunit med25 |
topic | Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4538835/ https://www.ncbi.nlm.nih.gov/pubmed/26130716 http://dx.doi.org/10.1093/nar/gkv650 |
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