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

Jean‐Philippe ‐P GALONS, Isei TANIDA, Yoshikazu OHYA, Yasuhiro ANRAKU, Yoji ARATA

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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
DOIs
StatePublished - Oct 1990
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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