Cargando…
A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae
Chemical sensitivity, growth inhibition in response to a chemical, is a powerful phenotype that can reveal insight into diverse cellular processes. Chemical sensitivity assays are used in nearly every model system, however the yeast Saccharomyces cerevisiae provides a particularly powerful platform...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6085351/ https://www.ncbi.nlm.nih.gov/pubmed/30093662 http://dx.doi.org/10.1038/s41598-018-30305-z |
_version_ | 1783346307968008192 |
---|---|
author | Hung, Chao-Wei Martínez-Márquez, Jorge Y. Javed, Fatima T. Duncan, Mara C. |
author_facet | Hung, Chao-Wei Martínez-Márquez, Jorge Y. Javed, Fatima T. Duncan, Mara C. |
author_sort | Hung, Chao-Wei |
collection | PubMed |
description | Chemical sensitivity, growth inhibition in response to a chemical, is a powerful phenotype that can reveal insight into diverse cellular processes. Chemical sensitivity assays are used in nearly every model system, however the yeast Saccharomyces cerevisiae provides a particularly powerful platform for discovery and mechanistic insight from chemical sensitivity assays. Here we describe a simple and inexpensive approach to determine chemical sensitivity quantitatively in yeast in the form of half maximal inhibitory concentration (IC(50)) using common laboratory equipment. We demonstrate the utility of this method using chemicals commonly used to monitor changes in membrane traffic. When compared to traditional agar-based plating methods, this method is more sensitive and can detect defects not apparent using other protocols. Additionally, this method reduces the experimental protocol from five days to 18 hours for the toxic amino acid canavanine. Furthermore, this method provides reliable results using lower amounts of chemicals. Finally, this method is easily adapted to additional chemicals as demonstrated with an engineered system that activates the spindle assembly checkpoint in response to rapamycin with differing efficiencies. This approach provides researchers with a cost-effective method to perform chemical genetic profiling without specialized equipment. |
format | Online Article Text |
id | pubmed-6085351 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-60853512018-08-16 A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae Hung, Chao-Wei Martínez-Márquez, Jorge Y. Javed, Fatima T. Duncan, Mara C. Sci Rep Article Chemical sensitivity, growth inhibition in response to a chemical, is a powerful phenotype that can reveal insight into diverse cellular processes. Chemical sensitivity assays are used in nearly every model system, however the yeast Saccharomyces cerevisiae provides a particularly powerful platform for discovery and mechanistic insight from chemical sensitivity assays. Here we describe a simple and inexpensive approach to determine chemical sensitivity quantitatively in yeast in the form of half maximal inhibitory concentration (IC(50)) using common laboratory equipment. We demonstrate the utility of this method using chemicals commonly used to monitor changes in membrane traffic. When compared to traditional agar-based plating methods, this method is more sensitive and can detect defects not apparent using other protocols. Additionally, this method reduces the experimental protocol from five days to 18 hours for the toxic amino acid canavanine. Furthermore, this method provides reliable results using lower amounts of chemicals. Finally, this method is easily adapted to additional chemicals as demonstrated with an engineered system that activates the spindle assembly checkpoint in response to rapamycin with differing efficiencies. This approach provides researchers with a cost-effective method to perform chemical genetic profiling without specialized equipment. Nature Publishing Group UK 2018-08-09 /pmc/articles/PMC6085351/ /pubmed/30093662 http://dx.doi.org/10.1038/s41598-018-30305-z Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Hung, Chao-Wei Martínez-Márquez, Jorge Y. Javed, Fatima T. Duncan, Mara C. A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae |
title | A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae |
title_full | A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae |
title_fullStr | A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae |
title_full_unstemmed | A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae |
title_short | A simple and inexpensive quantitative technique for determining chemical sensitivity in Saccharomyces cerevisiae |
title_sort | simple and inexpensive quantitative technique for determining chemical sensitivity in saccharomyces cerevisiae |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6085351/ https://www.ncbi.nlm.nih.gov/pubmed/30093662 http://dx.doi.org/10.1038/s41598-018-30305-z |
work_keys_str_mv | AT hungchaowei asimpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT martinezmarquezjorgey asimpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT javedfatimat asimpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT duncanmarac asimpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT hungchaowei simpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT martinezmarquezjorgey simpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT javedfatimat simpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae AT duncanmarac simpleandinexpensivequantitativetechniquefordeterminingchemicalsensitivityinsaccharomycescerevisiae |