Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters.

Détails

Ressource 1Télécharger: ncomms11304.pdf (2861.00 [Ko])
Etat: Public
Version: Final published version
ID Serval
serval:BIB_3F1CD4C83601
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters.
Périodique
Nature Communications
Auteur⸱e⸱s
Aymoz D., Wosika V., Durandau E., Pelet S.
ISSN
2041-1723 (Electronic)
ISSN-L
2041-1723
Statut éditorial
Publié
Date de publication
2016
Peer-reviewed
Oui
Volume
7
Pages
11304
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Résumé
Protein expression is a dynamic process, which can be rapidly induced by extracellular signals. It is widely appreciated that single cells can display large variations in the level of gene induction. However, the variability in the dynamics of this process in individual cells is difficult to quantify using standard fluorescent protein (FP) expression assays, due to the slow maturation of their fluorophore. Here we have developed expression reporters that accurately measure both the levels and dynamics of protein synthesis in live single cells with a temporal resolution under a minute. Our system relies on the quantification of the translocation of a constitutively expressed FP into the nucleus. As a proof of concept, we used these reporters to measure the transient protein synthesis arising from two promoters responding to the yeast hyper osmolarity glycerol mitogen-activated protein kinase pathway (pSTL1 and pGPD1). They display distinct expression dynamics giving rise to strikingly different instantaneous expression noise.
Mots-clé
Gene Expression Regulation, Fungal, Genes, Reporter, Glycerol/metabolism, Glycerol-3-Phosphate Dehydrogenase (NAD+)/genetics, Glycerol-3-Phosphate Dehydrogenase (NAD+)/metabolism, Green Fluorescent Proteins/genetics, Green Fluorescent Proteins/metabolism, Luminescent Proteins/genetics, Luminescent Proteins/metabolism, Membrane Transport Proteins/genetics, Membrane Transport Proteins/metabolism, Protein Biosynthesis, Protein Processing, Post-Translational, Protein Transport, Saccharomyces cerevisiae/genetics, Saccharomyces cerevisiae/metabolism, Saccharomyces cerevisiae Proteins/genetics, Saccharomyces cerevisiae Proteins/metabolism, Single-Cell Analysis/methods, Time Factors
Pubmed
Web of science
Open Access
Oui
Création de la notice
10/05/2016 18:55
Dernière modification de la notice
20/08/2019 14:36
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