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

Details

Ressource 1Download: ncomms11304.pdf (2861.00 [Ko])
State: Public
Version: Final published version
Serval ID
serval:BIB_3F1CD4C83601
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters.
Journal
Nature Communications
Author(s)
Aymoz D., Wosika V., Durandau E., Pelet S.
ISSN
2041-1723 (Electronic)
ISSN-L
2041-1723
Publication state
Published
Issued date
2016
Peer-reviewed
Oui
Volume
7
Pages
11304
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
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.
Keywords
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
Yes
Create date
10/05/2016 17:55
Last modification date
20/08/2019 13:36
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