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Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast

A yeast strain lacking Met4p, the primary transcriptional regulator of the sulfur assimilation pathway, cannot synthesize methionine. This apparently simple auxotroph did not grow well in rich media containing excess methionine, forming small colonies on yeast extract/peptone/dextrose plates. Faster...

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Autores principales: Hickman, Mark J., Petti, Allegra A., Ho-Shing, Olivia, Silverman, Sanford J., McIsaac, R. Scott, Lee, Traci A., Botstein, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3204079/
https://www.ncbi.nlm.nih.gov/pubmed/21900497
http://dx.doi.org/10.1091/mbc.E11-05-0467
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author Hickman, Mark J.
Petti, Allegra A.
Ho-Shing, Olivia
Silverman, Sanford J.
McIsaac, R. Scott
Lee, Traci A.
Botstein, David
author_facet Hickman, Mark J.
Petti, Allegra A.
Ho-Shing, Olivia
Silverman, Sanford J.
McIsaac, R. Scott
Lee, Traci A.
Botstein, David
author_sort Hickman, Mark J.
collection PubMed
description A yeast strain lacking Met4p, the primary transcriptional regulator of the sulfur assimilation pathway, cannot synthesize methionine. This apparently simple auxotroph did not grow well in rich media containing excess methionine, forming small colonies on yeast extract/peptone/dextrose plates. Faster-growing large colonies were abundant when overnight cultures were plated, suggesting that spontaneous suppressors of the growth defect arise with high frequency. To identify the suppressor mutations, we used genome-wide single-nucleotide polymorphism and standard genetic analyses. The most common suppressors were loss-of-function mutations in OPI1, encoding a transcriptional repressor of phospholipid metabolism. Using a new system that allows rapid and specific degradation of Met4p, we could study the dynamic expression of all genes following loss of Met4p. Experiments using this system with and without Opi1p showed that Met4 activates and Opi1p represses genes that maintain levels of S-adenosylmethionine (SAM), the substrate for most methyltransferase reactions. Cells lacking Met4p grow normally when either SAM is added to the media or one of the SAM synthetase genes is overexpressed. SAM is used as a methyl donor in three Opi1p-regulated reactions to create the abundant membrane phospholipid, phosphatidylcholine. Our results show that rapidly growing cells require significant methylation, likely for the biosynthesis of phospholipids.
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spelling pubmed-32040792012-01-16 Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast Hickman, Mark J. Petti, Allegra A. Ho-Shing, Olivia Silverman, Sanford J. McIsaac, R. Scott Lee, Traci A. Botstein, David Mol Biol Cell Articles A yeast strain lacking Met4p, the primary transcriptional regulator of the sulfur assimilation pathway, cannot synthesize methionine. This apparently simple auxotroph did not grow well in rich media containing excess methionine, forming small colonies on yeast extract/peptone/dextrose plates. Faster-growing large colonies were abundant when overnight cultures were plated, suggesting that spontaneous suppressors of the growth defect arise with high frequency. To identify the suppressor mutations, we used genome-wide single-nucleotide polymorphism and standard genetic analyses. The most common suppressors were loss-of-function mutations in OPI1, encoding a transcriptional repressor of phospholipid metabolism. Using a new system that allows rapid and specific degradation of Met4p, we could study the dynamic expression of all genes following loss of Met4p. Experiments using this system with and without Opi1p showed that Met4 activates and Opi1p represses genes that maintain levels of S-adenosylmethionine (SAM), the substrate for most methyltransferase reactions. Cells lacking Met4p grow normally when either SAM is added to the media or one of the SAM synthetase genes is overexpressed. SAM is used as a methyl donor in three Opi1p-regulated reactions to create the abundant membrane phospholipid, phosphatidylcholine. Our results show that rapidly growing cells require significant methylation, likely for the biosynthesis of phospholipids. The American Society for Cell Biology 2011-11-01 /pmc/articles/PMC3204079/ /pubmed/21900497 http://dx.doi.org/10.1091/mbc.E11-05-0467 Text en © 2011 Hickman et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Hickman, Mark J.
Petti, Allegra A.
Ho-Shing, Olivia
Silverman, Sanford J.
McIsaac, R. Scott
Lee, Traci A.
Botstein, David
Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
title Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
title_full Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
title_fullStr Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
title_full_unstemmed Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
title_short Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
title_sort coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3204079/
https://www.ncbi.nlm.nih.gov/pubmed/21900497
http://dx.doi.org/10.1091/mbc.E11-05-0467
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