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

Mark J. Hickman, Allegra A. Petti, Olivia Ho-Shing, Sanford J. Silverman, R. Scott McIsaac, Traci A. Lee, David Botstein

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

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 Opi1pregulated reactions to create the abundant membrane phospholipid, phosphatidylcholine. Our results show that rapidly growing cells require significant methylation, likely for the biosynthesis of phospholipids.

Original languageEnglish
Pages (from-to)4192-4204
Number of pages13
JournalMolecular biology of the cell
Volume22
Issue number21
DOIs
StatePublished - Nov 1 2011
Externally publishedYes

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