Discovery of lipid enzymes and their modulators using metabolite profiling of yeast (saccharomyces cerevisiae) mutants

Abstract

Identification of novel genes involved in the metabolism and regulation of lipids would allow for a greater insight into lipid metabolism. Comparative lipid profiling using mass spectrometry, whereby the lipid profile of a mutant and wild type strain are compared is one possible way to identify gene function. A total of one hundred and twenty mutant strains were profiled both randomly and on the basis coding for lipid related domains. Changes greater than the intra replicate variation were noted. Evidence for a possible role in glycerophospholipid metabolism was found for the ORF YBR042C. Deletion of this gene resulted in a specific decrease in phosphoinositol species. The sub cellular location of this gene was independently confirmed. In order to investigate substrate specificity a double knockout with SLC1 was carried out. Evidence linking an ORF to lipid metabolism was obtained and strategies to scale up the process are discussed.