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A new dominant peroxiredoxin allele identified by whole-genome re-sequencing of random mutagenized yeast causes oxidant-resistance and premature aging
The combination of functional genomics with next generation sequencing facilitates new experimental strategies for addressing complex biological phenomena. Here, we report the identification of a gain-of-function allele of peroxiredoxin (thioredoxin peroxidase, Tsa1p) via whole-genome re-sequencing...
Autores principales: | , , , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Impact Journals LLC
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2954039/ https://www.ncbi.nlm.nih.gov/pubmed/20729566 |
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author | Timmermann, Bernd Jarolim, Stefanie Rußmayer, Hannes Kerick, Martin Michel, Steve Krüger, Antje Bluemlein, Katharina Laun, Peter Grillari, Johannes Lehrach, Hans Breitenbach, Michael Ralser, Markus |
author_facet | Timmermann, Bernd Jarolim, Stefanie Rußmayer, Hannes Kerick, Martin Michel, Steve Krüger, Antje Bluemlein, Katharina Laun, Peter Grillari, Johannes Lehrach, Hans Breitenbach, Michael Ralser, Markus |
author_sort | Timmermann, Bernd |
collection | PubMed |
description | The combination of functional genomics with next generation sequencing facilitates new experimental strategies for addressing complex biological phenomena. Here, we report the identification of a gain-of-function allele of peroxiredoxin (thioredoxin peroxidase, Tsa1p) via whole-genome re-sequencing of a dominantSaccharomyces cerevisiae mutant obtained by chemical mutagenesis. Yeast strain K6001, a screening system for lifespan phenotypes, was treated with ethyl methanesulfonate (EMS). We isolated an oxidative stress-resistant mutant (B7) which transmitted this phenotype in a background-independent, monogenic and dominant way. By massive parallel pyrosequencing, we generated an 38.8 fold whole-genome coverage of the strains, which differed in 12,482 positions from the reference (S288c) genome. Via a subtraction strategy, we could narrow this number to 13 total and 4 missense nucleotide variations that were specific for the mutant. Via expression in wild type backgrounds, we show that one of these mutations, exchanging a residue in the peroxiredoxin Tsa1p, was responsible for the mutant phenotype causing background-independent dominant oxidative stress-resistance. These effects were not provoked by altered Tsa1p levels, nor could they be simulated by deletion, haploinsufficiency or over-expression of the wild-type allele. Furthermore, via both a mother-enrichment technique and a micromanipulation assay, we found a robust premature aging phenotype of this oxidant-resistant strain. Thus, TSA1-B7 encodes for a novel dominant form of peroxiredoxin, and establishes a new connection between oxidative stress and aging. In addition, this study shows that the re-sequencing of entire genomes is becoming a promising alternative for the identification of functional alleles in approaches of classic molecular genetics. |
format | Text |
id | pubmed-2954039 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-29540392010-10-14 A new dominant peroxiredoxin allele identified by whole-genome re-sequencing of random mutagenized yeast causes oxidant-resistance and premature aging Timmermann, Bernd Jarolim, Stefanie Rußmayer, Hannes Kerick, Martin Michel, Steve Krüger, Antje Bluemlein, Katharina Laun, Peter Grillari, Johannes Lehrach, Hans Breitenbach, Michael Ralser, Markus Aging (Albany NY) Research Article The combination of functional genomics with next generation sequencing facilitates new experimental strategies for addressing complex biological phenomena. Here, we report the identification of a gain-of-function allele of peroxiredoxin (thioredoxin peroxidase, Tsa1p) via whole-genome re-sequencing of a dominantSaccharomyces cerevisiae mutant obtained by chemical mutagenesis. Yeast strain K6001, a screening system for lifespan phenotypes, was treated with ethyl methanesulfonate (EMS). We isolated an oxidative stress-resistant mutant (B7) which transmitted this phenotype in a background-independent, monogenic and dominant way. By massive parallel pyrosequencing, we generated an 38.8 fold whole-genome coverage of the strains, which differed in 12,482 positions from the reference (S288c) genome. Via a subtraction strategy, we could narrow this number to 13 total and 4 missense nucleotide variations that were specific for the mutant. Via expression in wild type backgrounds, we show that one of these mutations, exchanging a residue in the peroxiredoxin Tsa1p, was responsible for the mutant phenotype causing background-independent dominant oxidative stress-resistance. These effects were not provoked by altered Tsa1p levels, nor could they be simulated by deletion, haploinsufficiency or over-expression of the wild-type allele. Furthermore, via both a mother-enrichment technique and a micromanipulation assay, we found a robust premature aging phenotype of this oxidant-resistant strain. Thus, TSA1-B7 encodes for a novel dominant form of peroxiredoxin, and establishes a new connection between oxidative stress and aging. In addition, this study shows that the re-sequencing of entire genomes is becoming a promising alternative for the identification of functional alleles in approaches of classic molecular genetics. Impact Journals LLC 2010-08-13 /pmc/articles/PMC2954039/ /pubmed/20729566 Text en Copyright: ©2010 Timmermann et al. http://creativecommons.org/licenses/by/2.5/ 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 work is properly cited. |
spellingShingle | Research Article Timmermann, Bernd Jarolim, Stefanie Rußmayer, Hannes Kerick, Martin Michel, Steve Krüger, Antje Bluemlein, Katharina Laun, Peter Grillari, Johannes Lehrach, Hans Breitenbach, Michael Ralser, Markus A new dominant peroxiredoxin allele identified by whole-genome re-sequencing of random mutagenized yeast causes oxidant-resistance and premature aging |
title | A new dominant peroxiredoxin allele identified by whole-genome
re-sequencing of random mutagenized yeast causes oxidant-resistance and
premature aging |
title_full | A new dominant peroxiredoxin allele identified by whole-genome
re-sequencing of random mutagenized yeast causes oxidant-resistance and
premature aging |
title_fullStr | A new dominant peroxiredoxin allele identified by whole-genome
re-sequencing of random mutagenized yeast causes oxidant-resistance and
premature aging |
title_full_unstemmed | A new dominant peroxiredoxin allele identified by whole-genome
re-sequencing of random mutagenized yeast causes oxidant-resistance and
premature aging |
title_short | A new dominant peroxiredoxin allele identified by whole-genome
re-sequencing of random mutagenized yeast causes oxidant-resistance and
premature aging |
title_sort | new dominant peroxiredoxin allele identified by whole-genome
re-sequencing of random mutagenized yeast causes oxidant-resistance and
premature aging |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2954039/ https://www.ncbi.nlm.nih.gov/pubmed/20729566 |
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