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Nucleic Acids Res. 2015 Aug 18;43(14):7110-21. doi: 10.1093/nar/gkv650. Epub 2015 Jun 29.

Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25.

Author information

1
CNRS UMR 8576, Université de Lille, Parc CNRS de la Haute Borne, 50 avenue de Halley, B.P. 70478, 59658 Villeneuve d'Ascq Cedex, France isabelle.landrieu@univ-lille1.fr.
2
CNRS UMR 8576, Université de Lille, Parc CNRS de la Haute Borne, 50 avenue de Halley, B.P. 70478, 59658 Villeneuve d'Ascq Cedex, France.
3
CNRS UMR 8576, Université de Lille, Parc CNRS de la Haute Borne, 50 avenue de Halley, B.P. 70478, 59658 Villeneuve d'Ascq Cedex, France didier.monte@iri.univ-lille1.fr.

Abstract

The N-terminal acidic transactivation domain (TAD) of ERM/ETV5 (ERM38-68), 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.

PMID:
26130716
PMCID:
PMC4538835
DOI:
10.1093/nar/gkv650
[Indexed for MEDLINE]
Free PMC Article

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