title: REV, A BRET-Based Sensor of ERK Activity.
abstract: Networks of signaling molecules are activated in response to environmental changes. How are these signaling networks dynamically integrated in space and time to process particular information? To tackle this issue, biosensors of single signaling pathways have been engineered. Bioluminescence resonance energy transfer (BRET)-based biosensors have proven to be particularly efficient in that matter due to the high sensitivity of this technology to monitor protein-protein interactions or conformational changes in living cells. Extracellular signal-regulated kinases (ERK) are ubiquitously expressed and involved in many diverse cellular functions that might be encoded by the strength and spatio-temporal pattern of ERK activation. We developed a BRET-based sensor of ERK activity, called Rluc8-ERKsubstrate-Venus (REV). As expected, BRET changes of REV were correlated with ERK phosphorylation, which is required for its kinase activity. In neurons, the nature of the stimuli determines the strength, the location, or the moment of ERK activation, thus highlighting how acute modulation of ERK may encode the nature of initial stimulus to specify the consequences of this activation. This study provides evidence for suitability of REV as a new biosensor to address biological questions.
authors.affiliation: CNRS, UMR-5203, Institut de Génomique Fonctionnelle , Montpellier , France ; INSERM, U661 , Montpellier , France ; UMR-5203, Universités de Montpellier 1 & 2 , Montpellier , France ; Sino-France Laboratory for Drug Screening, Key Laboratory of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology , Wuhan , China.
authors.forename: Chanjuan
authors.initials: C
authors.lastname: Xu
authors.forename: Marion
authors.initials: M
authors.lastname: Peter
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bouquier
authors.forename: Vincent
authors.initials: V
authors.lastname: Ollendorff
authors.forename: Ignacio
authors.initials: I
authors.lastname: Villamil
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Liu
authors.forename: Laurent
authors.initials: L
authors.lastname: Fagni
authors.forename: Julie
authors.initials: J
authors.lastname: Perroy
keywords: BRET imaging; Rluc8-ERKsubstrate-Venus; bioluminescence resonance energy transfer; biosensor; extracellular signal-regulated kinases; fluorescence lifetime imaging microscopy; spatio-temporal signaling
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