Cargando…

Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae

Development of fluorescent proteins (FPs) enabled researchers to visualize protein localization and trafficking in living cells and organisms. The extended palette of available FPs allows simultaneous detection of multiple fluorescent fusion proteins. Importantly, FPs are originally derived from dif...

Descripción completa

Detalles Bibliográficos
Autores principales: Albakri, Maram B., Jiang, Yuwei, Genereaux, Julie, Lajoie, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: F1000 Research Limited 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6290977/
https://www.ncbi.nlm.nih.gov/pubmed/30631438
http://dx.doi.org/10.12688/f1000research.15829.2
_version_ 1783380182279651328
author Albakri, Maram B.
Jiang, Yuwei
Genereaux, Julie
Lajoie, Patrick
author_facet Albakri, Maram B.
Jiang, Yuwei
Genereaux, Julie
Lajoie, Patrick
author_sort Albakri, Maram B.
collection PubMed
description Development of fluorescent proteins (FPs) enabled researchers to visualize protein localization and trafficking in living cells and organisms. The extended palette of available FPs allows simultaneous detection of multiple fluorescent fusion proteins. Importantly, FPs are originally derived from different organisms from jelly fish to corals and each FP displays its own biophysical properties. Among these properties, the tendency of FPs to oligomerize inherently affects the behavior of its fusion partner. Here we employed the budding yeast Saccharomyces cerevisiae to determine the impact of the latest generation of red FPs on their binding partner. We used a yeast assay based on the aggregation and toxicity of misfolded polyQ expansion proteins linked to Huntington’s disease. Since polyQ aggregation and toxicity are highly dependent on the sequences flanking the polyQ region, polyQ expansions provide an ideal tool to assess the impact of FPs on their fusion partners. We found that unlike what is observed for green FP variants, yemRFP and yFusionRed-tagged polyQ expansions  show reduced toxicity.  However, polyQ expansions tagged with the bright synthetically engineered ymScarlet displayed severe polyQ toxicity. Our data indicate that ymScarlet might have significant advantages over the previous generation of red FPs for use in fluorescent fusions in yeast.
format Online
Article
Text
id pubmed-6290977
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher F1000 Research Limited
record_format MEDLINE/PubMed
spelling pubmed-62909772019-01-09 Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae Albakri, Maram B. Jiang, Yuwei Genereaux, Julie Lajoie, Patrick F1000Res Research Note Development of fluorescent proteins (FPs) enabled researchers to visualize protein localization and trafficking in living cells and organisms. The extended palette of available FPs allows simultaneous detection of multiple fluorescent fusion proteins. Importantly, FPs are originally derived from different organisms from jelly fish to corals and each FP displays its own biophysical properties. Among these properties, the tendency of FPs to oligomerize inherently affects the behavior of its fusion partner. Here we employed the budding yeast Saccharomyces cerevisiae to determine the impact of the latest generation of red FPs on their binding partner. We used a yeast assay based on the aggregation and toxicity of misfolded polyQ expansion proteins linked to Huntington’s disease. Since polyQ aggregation and toxicity are highly dependent on the sequences flanking the polyQ region, polyQ expansions provide an ideal tool to assess the impact of FPs on their fusion partners. We found that unlike what is observed for green FP variants, yemRFP and yFusionRed-tagged polyQ expansions  show reduced toxicity.  However, polyQ expansions tagged with the bright synthetically engineered ymScarlet displayed severe polyQ toxicity. Our data indicate that ymScarlet might have significant advantages over the previous generation of red FPs for use in fluorescent fusions in yeast. F1000 Research Limited 2018-11-26 /pmc/articles/PMC6290977/ /pubmed/30631438 http://dx.doi.org/10.12688/f1000research.15829.2 Text en Copyright: © 2018 Albakri MB et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Note
Albakri, Maram B.
Jiang, Yuwei
Genereaux, Julie
Lajoie, Patrick
Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae
title Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae
title_full Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae
title_fullStr Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae
title_full_unstemmed Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae
title_short Polyglutamine toxicity assays highlight the advantages of mScarlet for imaging in Saccharomyces cerevisiae
title_sort polyglutamine toxicity assays highlight the advantages of mscarlet for imaging in saccharomyces cerevisiae
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6290977/
https://www.ncbi.nlm.nih.gov/pubmed/30631438
http://dx.doi.org/10.12688/f1000research.15829.2
work_keys_str_mv AT albakrimaramb polyglutaminetoxicityassayshighlighttheadvantagesofmscarletforimaginginsaccharomycescerevisiae
AT jiangyuwei polyglutaminetoxicityassayshighlighttheadvantagesofmscarletforimaginginsaccharomycescerevisiae
AT genereauxjulie polyglutaminetoxicityassayshighlighttheadvantagesofmscarletforimaginginsaccharomycescerevisiae
AT lajoiepatrick polyglutaminetoxicityassayshighlighttheadvantagesofmscarletforimaginginsaccharomycescerevisiae