Tandem fluorescent protein timers for non-invasive relative protein lifetime measurement in plants

A - Papers appearing in refereed journals

Zhang, H., Linster, E., Gannon, L., Leemhuis, W., Rundle, C., Theodoulou, F. L. and Wirtz, M. 2019. Tandem fluorescent protein timers for non-invasive relative protein lifetime measurement in plants. Plant Physiology.

AuthorsZhang, H., Linster, E., Gannon, L., Leemhuis, W., Rundle, C., Theodoulou, F. L. and Wirtz, M.
Abstract

Targeted protein degradation is an important and pervasive regulatory mechanism in plants, required for perception and response to the environment as well as developmental signalling. Despite the significance of this process, relatively few studies have assessed plant protein turnover in a quantitative fashion. Tandem fluorescent protein timers (tFTs) offer a powerful approach for the assessment of in vivo protein turnover in distinct subcellular compartments of single or multiple cells. A tFT is a fusion of two different fluorescent proteins with distinct fluorophore maturation kinetics, which enable protein age to be estimated from the ratio of fluorescence intensities of the two fluorescent proteins. Here, we use short-lived auxin signalling proteins and model N-end rule pathway reporters to demonstrate the utility of tFTs for studying protein turnover in living plants. We present transient expression of tFTs as an efficient screen for relative protein lifetime, useful for testing the effects of mutations and different genetic backgrounds on protein stability, and demonstrate the potential for using stably expressed tFTs to study native protein dynamics with high temporal resolution in response to exogenous or endogenous stimuli.

KeywordsProtein lifetime; Proteostasis; Arabidopsi; Tobacco; Fluorescent protein
Year of Publication2019
JournalPlant Physiology
Digital Object Identifier (DOI)doi:10.1104/pp.19.00051
PubMed ID30872425
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeTPM - Tailoring Plant Metabolism - Work package 1 (WP1) - High value lipids for health and industry
Substrates of the N-end rule of targeted protein degradation
Accepted author manuscript
Output statusPublished
Publication dates
Online14 Mar 2019
Publication process dates
Accepted08 Mar 2019
PublisherAmerican Society of Plant Biologists (ASPB)
Copyright licenseCC BY
ISSN0032-0889

Permalink - https://repository.rothamsted.ac.uk/item/8w8w3/tandem-fluorescent-protein-timers-for-non-invasive-relative-protein-lifetime-measurement-in-plants

86 total views
105 total downloads
0 views this month
0 downloads this month
Download files as zip