Polyglutamine toxicity in yeast uncovers phenotypic variations between different fluorescent protein fusions. (1st November 2016)

• Record Type: Journal Article
• Authors: Jiang, Yuwei; Di Gregorio, Sonja E.; Duennwald, Martin L.; Lajoie, Patrick
• Is Part Of: Traffic. Volume 18:Number 1(2017)
• Page Start: 58
• Page End: 70
• Publication Date: 2016-11-01
• Subjects: fluorescent proteins -- Huntington's disease -- polyglutamine; Biological transport -- Periodicals; 571.6
• DOI: 10.1111/tra.12453
• Languages: English
• ISSNs: 1398-9219

Abstract

Abstract : The palette of fluorescent proteins (FPs) available for live‐cell imaging contains proteins that strongly differ in their biophysical properties. FPs cannot be assumed to be equivalent and in certain cases could significantly perturb the behavior of fluorescent reporters. We employed Saccharomyces cerevisiae to comprehensively study the impact of FPs on the toxicity of polyglutamine (polyQ) expansion proteins associated with Huntington's disease. The toxicity of polyQ fusion constructs is highly dependent on the sequences flanking the polyQ repeats. Thus, they represent a powerful tool to study the impact of fluorescent fusion partners. We observed significant differences on polyQ aggregation and toxicity between commonly used FPs. We generated a novel series of vectors with latest yeast‐optimized FPs for investigation of Htt toxicity, including a newly optimized blue FP for expression in yeast. Our study highlights the importance of carefully choosing the optimal FPs when designing tagging strategies. Abstract : Fluorescent proteins have revolutionized the work of scientists by allowing the direct visualization of protein trafficking and localization in living cells. However, fluorescent proteins are too often seen as interchangeable building blocks than can be easily swapped to accommodate a specific imaging condition. In this study, we used a yeast model of Huntington's diseases to highlight the differential impact of the most common fluorescent proteins on the aggregation and toxicity of expanded polyglutamine proteins.