title: Opposing effects of Elk-1 multisite phosphorylation shape its response to ERK activation.
abstract: Multisite phosphorylation regulates many transcription factors, including the serum response factor partner Elk-1. Phosphorylation of the transcriptional activation domain (TAD) of Elk-1 by the protein kinase ERK at multiple sites potentiates recruitment of the Mediator transcriptional coactivator complex and transcriptional activation, but the roles of individual phosphorylation events had remained unclear. Using time-resolved nuclear magnetic resonance spectroscopy, we found that ERK2 phosphorylation proceeds at markedly different rates at eight TAD sites in vitro, which we classified as fast, intermediate, and slow. Mutagenesis experiments showed that phosphorylation of fast and intermediate sites promoted Mediator interaction and transcriptional activation, whereas modification of slow sites counteracted both functions, thereby limiting Elk-1 output. Progressive Elk-1 phosphorylation thus ensures a self-limiting response to ERK activation, which occurs independently of antagonizing phosphatase activity.
authors.affiliation: Signalling and Transcription Laboratory, Francis Crick Institute, Lincoln's Inn Fields Laboratory, London WC2A 3LY, UK.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Mylona
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-Supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany.
authors.forename: Francois-Xavier
authors.initials: FX
authors.lastname: Theillet
authors.affiliation: Signalling and Transcription Laboratory, Francis Crick Institute, Lincoln's Inn Fields Laboratory, London WC2A 3LY, UK.
authors.forename: Charles
authors.initials: C
authors.lastname: Foster
authors.affiliation: Biomolecular Modelling Laboratory, Francis Crick Institute, Lincoln's Inn Fields Laboratory, London WC2A 3LY, UK.
authors.forename: Tammy M
authors.initials: TM
authors.lastname: Cheng
authors.affiliation: Signalling and Transcription Laboratory, Francis Crick Institute, Lincoln's Inn Fields Laboratory, London WC2A 3LY, UK. Molecular & Clinical Sciences Research Institute, St. George's, University of London, London SW17 0RE, UK.
authors.forename: Francesc
authors.initials: F
authors.lastname: Miralles
authors.affiliation: Biomolecular Modelling Laboratory, Francis Crick Institute, Lincoln's Inn Fields Laboratory, London WC2A 3LY, UK.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Bates
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-Supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Selenko
authors.affiliation: Signalling and Transcription Laboratory, Francis Crick Institute, Lincoln's Inn Fields Laboratory, London WC2A 3LY, UK. richard.treisman@crick.ac.uk.
authors.forename: Richard
authors.initials: R
authors.lastname: Treisman
mesh_terms: D000818:Animals; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D006801:Humans; D020935:MAP Kinase Signaling System; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D016296:Mutagenesis; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation; D000072417:Protein Domains; D026362:Serum Response Factor / Q000378:metabolism; D050807:ets-Domain Protein Elk-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D026362:Serum Response Factor; D050807:ets-Domain Protein Elk-1; D019950:Mitogen-Activated Protein Kinase 1