Toward a global analysis of metabolites in regulatory mutants of yeast

Elizabeth M. Humston, Kenneth M. Dombek, Benjamin P. Tu, Elton T. Young, Robert E. Synovec

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The AMP-activated protein kinase in yeast, Snf1, coordinates expression and activity of numerous intracellular signaling and developmental pathways, including those regulating cellular differentiation, response to stress, meiosis, autophagy, and the diauxic transition. Snf1 phosphorylates metabolic enzymes and transcription factors to change cellular physiology and metabolism. Adr1 and Cat8, transcription factors that activate gene expression after the diauxic transition, are regulated by Snf1; Cat8 through direct phosphorylation and Adr1 by dephosphorylation in a Snf1-dependent manner. Adr1 and Cat8 coordinately regulate numerous genes encoding enzymes of gluconeogenesis, the glyoxylate cycle, β-oxidation of fatty acids, and the utilization of alternative fermentable sugars and nonfermentable substrates. To determine the roles of Adr1, Cat8, and Snf1 in metabolism, two-dimensional gas chromatography coupled to time-of-flight mass spectrometry and liquid chromatography coupled to tandem mass spectrometry were used to identify metabolites whose levels change after the diauxic transition in wild-type-, ADR1-, CAT8-, and SNF1-deficient yeast. A discovery-based approach to data analysis utilized chemometric algorithms to identify, quantify, and compare 63 unique metabolites between wild type, adr1Δ, cat8Δ, adr1Δcat8Δ, and snf1Δ strains. The primary metabolites found to differ were those of gluconeogenesis, the glyoxylate and tricarboxylic acid cycles, and amino acid metabolism. In general, good agreement was observed between the levels of metabolites derived from these pathways and the levels of transcripts from the same strains, suggesting that transcriptional control plays a major role in regulating the levels of metabolites after the diauxic transition. [Figure not available: see fulltext.]

Original languageEnglish (US)
Pages (from-to)2387-2402
Number of pages16
JournalAnalytical and Bioanalytical Chemistry
Volume401
Issue number8
DOIs
StatePublished - Nov 2011

Fingerprint

Metabolites
Yeast
Gluconeogenesis
Yeasts
Metabolism
Transcription Factors
AMP-Activated Protein Kinases
Citric Acid Cycle
Autophagy
Meiosis
Mass spectrometry
Enzymes
Tandem Mass Spectrometry
Liquid Chromatography
Gas Chromatography
Mass Spectrometry
Fatty Acids
Phosphorylation
Gene encoding
Liquid chromatography

Keywords

  • Chemometrics
  • GC×GC-TOFMS
  • PARAFAC
  • Snf1 protein complex

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Toward a global analysis of metabolites in regulatory mutants of yeast. / Humston, Elizabeth M.; Dombek, Kenneth M.; Tu, Benjamin P.; Young, Elton T.; Synovec, Robert E.

In: Analytical and Bioanalytical Chemistry, Vol. 401, No. 8, 11.2011, p. 2387-2402.

Research output: Contribution to journalArticle

Humston, Elizabeth M. ; Dombek, Kenneth M. ; Tu, Benjamin P. ; Young, Elton T. ; Synovec, Robert E. / Toward a global analysis of metabolites in regulatory mutants of yeast. In: Analytical and Bioanalytical Chemistry. 2011 ; Vol. 401, No. 8. pp. 2387-2402.
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