A multinuclear magnetic resonance study of a cls11 mutant showing the Pet- phenotype of Saccharomyces cerevisiae

Jean-Philippe Galons, I. Tanida, Y. Ohya, Y. Anraku, Y. Arata

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2 Citations (Scopus)

Abstract

Energetic and intermediary metabolism was studied in a Pet- mutant of Saccharomyces cerevisiae with a calcium-sensitive phenotype that shows an inability to grow when cultured in a medium containing non-fermentable substrates. The perchloric acid extracts were prepared from suspensions of cls11 mutant and wild-type cells incubated with [1,3-13C]glycerol or [2-13C]acetate, and analyzed by 31P, 13C and 1H NMR. 31P- and 1H-NMR spectra showed significant differences between cls11 and wild-type cells at the level of amino acids, the storage carbohydrate trehalose (higher in mutant cells), and sugar phosphates (higher in wild-type cells). 13C-NMR spectra revealed major differences in the steady-state labelling of glutamate carbons. For incubations with [1,3-13C]glycerol, we estimated from the relative 13C enrichment of glutamate carbons that acetyl-CoA C2 is 43% C13 labelled in wild-type and 10% 13C labelled in mutant cells, respectively. For incubations with [2-13C]acetate, we calculated that the ratio of the relative flux through the glyoxylate shunt versus oxidative reactions is 58% in wild-type cells and 44% in the cls11 mutant cells. Again, a dilution of the relative enrichment of C2 of acetyl-CoA was observed in the mutant cells (89%) compared to the wild-type cells (97%). These results are discussed in terms of pleiotropic defects in non-fermentable carbon metabolism in mutant cells.

Original languageEnglish (US)
Pages (from-to)111-119
Number of pages9
JournalEuropean Journal of Biochemistry
Volume193
Issue number1
StatePublished - 1990
Externally publishedYes

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Pets
Magnetic resonance
Yeast
Saccharomyces cerevisiae
Acetyl Coenzyme A
Magnetic Resonance Spectroscopy
Carbon
Nuclear magnetic resonance
Phenotype
Metabolism
Glycerol
Glutamic Acid
Acetates
Sugar Phosphates
Trehalose
Labeling
Dilution
Suspensions
Carbohydrates
Fluxes

ASJC Scopus subject areas

  • Biochemistry

Cite this

A multinuclear magnetic resonance study of a cls11 mutant showing the Pet- phenotype of Saccharomyces cerevisiae. / Galons, Jean-Philippe; Tanida, I.; Ohya, Y.; Anraku, Y.; Arata, Y.

In: European Journal of Biochemistry, Vol. 193, No. 1, 1990, p. 111-119.

Research output: Contribution to journalArticle

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