Characterization of a Saccharomyces cerevisiae mutant defective in inositol sphingolipid synthesis
A mutant of S. cerevisiae, auxotrophic for the sphingolipid long chain bases sphinganine or 4-hydroxysphinganine has been further characterized. The mutant is defective in the first step in the synthesis of sphingolipid long chain bases, the formation of 3-keto-sphinganine from palmitoyl CoA and L-serine. Crude membranes from wild type cells exhibit a palmitoyl CoA dependent conversion of (/sup 3/H)serine to a labeled product with the properties of 3-ketosphinganine, whereas membranes from the mutant exhibit negligible activity. As expected from this result, they authors find that 3-ketosphinganine supports growth of the mutant as effectively as erythro-sphinganine. To study the sequelae of long chain base deprivation in the mutant required a method to effectively remove lipophilic long chain bases from the cultured cells. They have found that addition of ..cap alpha..-cyclodextrin to the culture medium evidently binds sphinganine tightly enough such that within an hour they observe inhibition of growth (turbidity, cell count) and incorporation of (/sup 3/H)inositol into sphingolipids. Such treatment also results in a loss of cell viability, a long chain base less death possibly similar to the inositol-less death observed in inositol auxotrophs of yeast.
- Research Organization:
- Univ. of Kentucky, Lexington
- OSTI ID:
- 7246738
- Report Number(s):
- CONF-8606151-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550701 -- Microbiology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
CARBOHYDRATES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL COMPOSITION
FUNGI
GROWTH
HYDROXY ACIDS
INOSITOL
INOSITOLS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LIPIDS
MEMBRANES
MICROORGANISMS
MONOSACCHARIDES
MUTANTS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
REACTION KINETICS
SACCHARIDES
SACCHAROMYCES
SACCHAROMYCES CEREVISIAE
SERINE
SYNTHESIS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VIABILITY
YEASTS