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Global analysis of protein homomerization in Saccharomyces cerevisiae

In vivo analyses of the occurrence, subcellular localization, and dynamics of protein–protein interactions (PPIs) are important issues in functional proteomic studies. The bimolecular fluorescence complementation (BiFC) assay has many advantages in that it provides a reliable way to detect PPIs in l...

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Detalles Bibliográficos
Autores principales: Kim, Yeonsoo, Jung, Jong Pil, Pack, Chan-Gi, Huh, Won-Ki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6314163/
https://www.ncbi.nlm.nih.gov/pubmed/30567710
http://dx.doi.org/10.1101/gr.231860.117
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author Kim, Yeonsoo
Jung, Jong Pil
Pack, Chan-Gi
Huh, Won-Ki
author_facet Kim, Yeonsoo
Jung, Jong Pil
Pack, Chan-Gi
Huh, Won-Ki
author_sort Kim, Yeonsoo
collection PubMed
description In vivo analyses of the occurrence, subcellular localization, and dynamics of protein–protein interactions (PPIs) are important issues in functional proteomic studies. The bimolecular fluorescence complementation (BiFC) assay has many advantages in that it provides a reliable way to detect PPIs in living cells with minimal perturbation of the structure and function of the target proteins. Previously, to facilitate the application of the BiFC assay to genome-wide analysis of PPIs, we generated a collection of yeast strains expressing full-length proteins tagged with the N-terminal fragment of Venus (VN), a yellow fluorescent protein variant, from their own native promoters. In the present study, we constructed a VC (the C-terminal fragment of Venus) fusion library consisting of 5671 MATα strains expressing C-terminally VC-tagged proteins (representing ∼91% of the yeast proteome). For genome-wide analysis of protein homomer formation, we mated each strain in the VC fusion library with its cognate strain in the VN fusion library and performed the BiFC assay. From this analysis, we identified 186 homomer candidates. We further investigated the functional relevance of the homomerization of Pln1, a yeast perilipin. Our data set provides a useful resource for understanding the physiological roles of protein homomerization. Furthermore, the VC fusion library together with the VN fusion library will provide a valuable platform to systematically analyze PPIs in the natural cellular context.
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spelling pubmed-63141632019-07-01 Global analysis of protein homomerization in Saccharomyces cerevisiae Kim, Yeonsoo Jung, Jong Pil Pack, Chan-Gi Huh, Won-Ki Genome Res Resource In vivo analyses of the occurrence, subcellular localization, and dynamics of protein–protein interactions (PPIs) are important issues in functional proteomic studies. The bimolecular fluorescence complementation (BiFC) assay has many advantages in that it provides a reliable way to detect PPIs in living cells with minimal perturbation of the structure and function of the target proteins. Previously, to facilitate the application of the BiFC assay to genome-wide analysis of PPIs, we generated a collection of yeast strains expressing full-length proteins tagged with the N-terminal fragment of Venus (VN), a yellow fluorescent protein variant, from their own native promoters. In the present study, we constructed a VC (the C-terminal fragment of Venus) fusion library consisting of 5671 MATα strains expressing C-terminally VC-tagged proteins (representing ∼91% of the yeast proteome). For genome-wide analysis of protein homomer formation, we mated each strain in the VC fusion library with its cognate strain in the VN fusion library and performed the BiFC assay. From this analysis, we identified 186 homomer candidates. We further investigated the functional relevance of the homomerization of Pln1, a yeast perilipin. Our data set provides a useful resource for understanding the physiological roles of protein homomerization. Furthermore, the VC fusion library together with the VN fusion library will provide a valuable platform to systematically analyze PPIs in the natural cellular context. Cold Spring Harbor Laboratory Press 2019-01 /pmc/articles/PMC6314163/ /pubmed/30567710 http://dx.doi.org/10.1101/gr.231860.117 Text en © 2019 Kim et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Resource
Kim, Yeonsoo
Jung, Jong Pil
Pack, Chan-Gi
Huh, Won-Ki
Global analysis of protein homomerization in Saccharomyces cerevisiae
title Global analysis of protein homomerization in Saccharomyces cerevisiae
title_full Global analysis of protein homomerization in Saccharomyces cerevisiae
title_fullStr Global analysis of protein homomerization in Saccharomyces cerevisiae
title_full_unstemmed Global analysis of protein homomerization in Saccharomyces cerevisiae
title_short Global analysis of protein homomerization in Saccharomyces cerevisiae
title_sort global analysis of protein homomerization in saccharomyces cerevisiae
topic Resource
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6314163/
https://www.ncbi.nlm.nih.gov/pubmed/30567710
http://dx.doi.org/10.1101/gr.231860.117
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