Tandem fluorescent protein timers for noninvasive relative protein lifetime measurement in plants
<p>Targeted protein degradation is an important and pervasive regulatory mechanism in plants, required for perception and response to the environment as well as developmental signaling. Despite the significance of this process, relatively few studies have assessed plant protein turnover in a q...
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| Main Authors: | , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
June 2019
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| In: |
Plant physiology
Year: 2019, Volume: 180, Issue: 2, Pages: 718-731 |
| ISSN: | 1532-2548 |
| DOI: | 10.1104/pp.19.00051 |
| Online Access: | Verlag, Volltext: https://doi.org/10.1104/pp.19.00051 Verlag, Volltext: http://www.plantphysiol.org/content/180/2/718 
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| Author Notes: | Hongtao Zhang, Eric Linster, Lucy Gannon, Wiebke Leemhuis, Chelsea A. Rundle, Frederica L. Theodoulou, and Markus Wirtz |
| Summary: | <p>Targeted protein degradation is an important and pervasive regulatory mechanism in plants, required for perception and response to the environment as well as developmental signaling. 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 used short-lived auxin signaling proteins and model N-end rule (N-recognin) pathway reporters to demonstrate the utility of tFTs for studying protein turnover in living plant cells of Arabidopsis (<i>Arabidopsis thaliana</i>) and <i>Nicotiana benthamiana</i>. 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. This work demonstrates the potential for using stably expressed tFTs to study native protein dynamics with high temporal resolution in response to exogenous or endogenous stimuli.</p> |
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| Item Description: | Gesehen am 17.09.2019 |
| Physical Description: | Online Resource |
| ISSN: | 1532-2548 |
| DOI: | 10.1104/pp.19.00051 |