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Rapid identification of mRNA processing defects with a novel single-cell yeast reporter
It has become increasingly evident that gene expression processes in eukaryotes involve communication and coordination between many complex, independent macromolecular machines. To query these processes and to explore the potential relationships between them in the budding yeast Saccharomyces cerevi...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3988574/ https://www.ncbi.nlm.nih.gov/pubmed/24671766 http://dx.doi.org/10.1261/rna.042663.113 |
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author | Sorenson, Matthew R. Stevens, Scott W. |
author_facet | Sorenson, Matthew R. Stevens, Scott W. |
author_sort | Sorenson, Matthew R. |
collection | PubMed |
description | It has become increasingly evident that gene expression processes in eukaryotes involve communication and coordination between many complex, independent macromolecular machines. To query these processes and to explore the potential relationships between them in the budding yeast Saccharomyces cerevisiae, we designed a versatile reporter using multicolor high-throughput flow cytometry. Due to its design, this single reporter exhibits a distinctive signature for many defects in gene expression including transcription, histone modification, pre-mRNA splicing, mRNA export, nonsense-mediated decay, and mRNA degradation. Analysis of the reporter in 4967 nonessential yeast genes revealed striking phenotypic overlaps between chromatin remodeling, histone modification, and pre-mRNA splicing. Additionally, we developed a copper-inducible reporter, with which we demonstrate that 5-fluorouracil mimics the mRNA decay phenotype of cells lacking the 3′–5′ exonuclease Rrp6p. Our reporter is capable of performing high-throughput, rapid, and large-scale screens to identify and characterize genetic and chemical perturbations of the major eukaryotic gene expression processes. |
format | Online Article Text |
id | pubmed-3988574 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-39885742015-05-01 Rapid identification of mRNA processing defects with a novel single-cell yeast reporter Sorenson, Matthew R. Stevens, Scott W. RNA Methods It has become increasingly evident that gene expression processes in eukaryotes involve communication and coordination between many complex, independent macromolecular machines. To query these processes and to explore the potential relationships between them in the budding yeast Saccharomyces cerevisiae, we designed a versatile reporter using multicolor high-throughput flow cytometry. Due to its design, this single reporter exhibits a distinctive signature for many defects in gene expression including transcription, histone modification, pre-mRNA splicing, mRNA export, nonsense-mediated decay, and mRNA degradation. Analysis of the reporter in 4967 nonessential yeast genes revealed striking phenotypic overlaps between chromatin remodeling, histone modification, and pre-mRNA splicing. Additionally, we developed a copper-inducible reporter, with which we demonstrate that 5-fluorouracil mimics the mRNA decay phenotype of cells lacking the 3′–5′ exonuclease Rrp6p. Our reporter is capable of performing high-throughput, rapid, and large-scale screens to identify and characterize genetic and chemical perturbations of the major eukaryotic gene expression processes. Cold Spring Harbor Laboratory Press 2014-05 /pmc/articles/PMC3988574/ /pubmed/24671766 http://dx.doi.org/10.1261/rna.042663.113 Text en © 2014 Sorenson and Stevens; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 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 | Methods Sorenson, Matthew R. Stevens, Scott W. Rapid identification of mRNA processing defects with a novel single-cell yeast reporter |
title | Rapid identification of mRNA processing defects with a novel single-cell yeast reporter |
title_full | Rapid identification of mRNA processing defects with a novel single-cell yeast reporter |
title_fullStr | Rapid identification of mRNA processing defects with a novel single-cell yeast reporter |
title_full_unstemmed | Rapid identification of mRNA processing defects with a novel single-cell yeast reporter |
title_short | Rapid identification of mRNA processing defects with a novel single-cell yeast reporter |
title_sort | rapid identification of mrna processing defects with a novel single-cell yeast reporter |
topic | Methods |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3988574/ https://www.ncbi.nlm.nih.gov/pubmed/24671766 http://dx.doi.org/10.1261/rna.042663.113 |
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