title: Coordinated regulation of sulfur and phospholipid metabolism reflects the importance of methylation in the growth of yeast.
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 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.
authors: Lewis-Sigler Institute for Integrative Genomics and Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA. hickmanm@tcnj.edu
authors: Mark J
authors: MJ
authors: Hickman
authors: Allegra A
authors: AA
authors: Petti
authors: Olivia
authors: O
authors: Ho-Shing
authors: Sanford J
authors: SJ
authors: Silverman
authors: R Scott
authors: RS
authors: McIsaac
authors: Traci A
authors: TA
authors: Lee
authors: David
authors: D
authors: Botstein
mesh_terms: D019943:Amino Acid Substitution; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D055786:Gene Knockout Techniques; D008715:Methionine / Q000378:metabolism; D008745:Methylation; D009154:Mutation; D007296:Myo-Inositol-1-Phosphate Synthase / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010743:Phospholipids / Q000096:biosynthesis / Q000378:metabolism*; D020641:Polymorphism, Single Nucleotide; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; C073675:MET4 protein, S cerevisiae; D009687:Nuclear Proteins; C096105:OPI1 protein, S cerevisiae; D010743:Phospholipids; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C066502:TUP1 protein, S cerevisiae; D013455:Sulfur; D012436:S-Adenosylmethionine; D008715:Methionine; C519641:INO1 protein, S cerevisiae; D007296:Myo-Inositol-1-Phosphate Synthase