Cargando…
Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe
Synthetic genetic array (SGA) has been successfully used to identify genetic interactions in S. cerevisiae and S. pombe. In S. pombe, SGA methods use either cycloheximide (C) or heat shock (HS) to select double mutants before measuring colony size as a surrogate for fitness. Quantitative Fitness Ana...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4500466/ https://www.ncbi.nlm.nih.gov/pubmed/26168240 http://dx.doi.org/10.1371/journal.pone.0132240 |
_version_ | 1782380919271718912 |
---|---|
author | Narayanan, Siddharth Dubarry, Marion Lawless, Conor Banks, A. Peter Wilkinson, Darren J. Whitehall, Simon K. Lydall, David |
author_facet | Narayanan, Siddharth Dubarry, Marion Lawless, Conor Banks, A. Peter Wilkinson, Darren J. Whitehall, Simon K. Lydall, David |
author_sort | Narayanan, Siddharth |
collection | PubMed |
description | Synthetic genetic array (SGA) has been successfully used to identify genetic interactions in S. cerevisiae and S. pombe. In S. pombe, SGA methods use either cycloheximide (C) or heat shock (HS) to select double mutants before measuring colony size as a surrogate for fitness. Quantitative Fitness Analysis (QFA) is a different method for determining fitness of microbial strains. In QFA, liquid cultures are spotted onto solid agar and growth curves determined for each spot by photography and model fitting. Here, we compared the two S. pombe SGA methods and found that the HS method was more reproducible for us. We also developed a QFA procedure for S. pombe. We used QFA to identify genetic interactions affecting two temperature sensitive, telomere associated query mutations (taz1Δ and pot1-1). We identify exo1∆ and other gene deletions as suppressors or enhancers of S. pombe telomere defects. Our study identifies known and novel gene deletions affecting the fitness of strains with telomere defects. The interactions we identify may be relevant in human cells. |
format | Online Article Text |
id | pubmed-4500466 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45004662015-07-17 Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe Narayanan, Siddharth Dubarry, Marion Lawless, Conor Banks, A. Peter Wilkinson, Darren J. Whitehall, Simon K. Lydall, David PLoS One Research Article Synthetic genetic array (SGA) has been successfully used to identify genetic interactions in S. cerevisiae and S. pombe. In S. pombe, SGA methods use either cycloheximide (C) or heat shock (HS) to select double mutants before measuring colony size as a surrogate for fitness. Quantitative Fitness Analysis (QFA) is a different method for determining fitness of microbial strains. In QFA, liquid cultures are spotted onto solid agar and growth curves determined for each spot by photography and model fitting. Here, we compared the two S. pombe SGA methods and found that the HS method was more reproducible for us. We also developed a QFA procedure for S. pombe. We used QFA to identify genetic interactions affecting two temperature sensitive, telomere associated query mutations (taz1Δ and pot1-1). We identify exo1∆ and other gene deletions as suppressors or enhancers of S. pombe telomere defects. Our study identifies known and novel gene deletions affecting the fitness of strains with telomere defects. The interactions we identify may be relevant in human cells. Public Library of Science 2015-07-13 /pmc/articles/PMC4500466/ /pubmed/26168240 http://dx.doi.org/10.1371/journal.pone.0132240 Text en © 2015 Narayanan et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Narayanan, Siddharth Dubarry, Marion Lawless, Conor Banks, A. Peter Wilkinson, Darren J. Whitehall, Simon K. Lydall, David Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe |
title | Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe
|
title_full | Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe
|
title_fullStr | Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe
|
title_full_unstemmed | Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe
|
title_short | Quantitative Fitness Analysis Identifies exo1∆ and Other Suppressors or Enhancers of Telomere Defects in Schizosaccharomyces pombe
|
title_sort | quantitative fitness analysis identifies exo1∆ and other suppressors or enhancers of telomere defects in schizosaccharomyces pombe |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4500466/ https://www.ncbi.nlm.nih.gov/pubmed/26168240 http://dx.doi.org/10.1371/journal.pone.0132240 |
work_keys_str_mv | AT narayanansiddharth quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe AT dubarrymarion quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe AT lawlessconor quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe AT banksapeter quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe AT wilkinsondarrenj quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe AT whitehallsimonk quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe AT lydalldavid quantitativefitnessanalysisidentifiesexo1andothersuppressorsorenhancersoftelomeredefectsinschizosaccharomycespombe |