title: Polyglutamine toxicity in yeast uncovers phenotypic variations between different fluorescent protein fusions.
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.
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, Canada.
authors.forename: Yuwei
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Department of Pathology and Laboratory Medicine, The University of Western Ontario, London, Canada.
authors.forename: Sonja E
authors.initials: SE
authors.lastname: Di Gregorio
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, Canada.|Department of Pathology and Laboratory Medicine, The University of Western Ontario, London, Canada.
authors.forename: Martin L
authors.initials: ML
authors.lastname: Duennwald
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, Canada.
authors.forename: Patrick
authors.initials: P
authors.lastname: Lajoie
mesh_terms: D006801:Humans; D006816:Huntington Disease / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: D008164:Luminescent Proteins; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; C097188:polyglutamine
keywords: Huntington's disease; fluorescent proteins; polyglutamine