Fatty acid desaturase mutants of yeast: growth requirements and electron spin resonance spin-label distribution

Abstract

Two respiratory-sufficient and one respiratory-deficient (nuclear petite) strains of yeast Delta(9)-desaturase mutants were analyzed to determine which fatty acids would serve as replacements for the naturally occurring fatty acids, 16:1 Delta(9)cis and 18:1 Delta(9)cis. The requirement can be satisfied by several fatty acids differing in double-bond position, steric configuration, chain length, and degree of unsaturation. The features common to growth-supporting fatty acids are presented and the effects of varying the carbon source and temperature are considered. In addition, we illustrate several pitfalls encountered in membrane studies which exploit lipid-requiring organisms. Since the membrane fatty acid composition of these mutants can be modified readily, electron spin resonance spectroscopy is used to compare membranes of mutant strains enriched for different fatty acids. The lipid distribution pattern of the most commonly employed electron spin resonance spin-label, 12-nitroxide stearate, was ascertained and compared to that of 18:1 Delta(9)cis.