title: High-content screening of yeast mutant libraries by shotgun lipidomics.
abstract: To identify proteins with a functional role in lipid metabolism and homeostasis we designed a high-throughput platform for high-content lipidomic screening of yeast mutant libraries. To this end, we combined culturing and lipid extraction in 96-well format, automated direct infusion nanoelectrospray ionization, high-resolution Orbitrap mass spectrometry, and a dedicated data processing framework to support lipid phenotyping across hundreds of Saccharomyces cerevisiae mutants. Our novel approach revealed that the absence of genes with unknown function YBR141C and YJR015W, and the transcription factor KAR4 precipitated distinct lipid metabolic phenotypes. These results demonstrate that the high-throughput shotgun lipidomics platform is a valid and complementary proxy for high-content screening of yeast mutant libraries.
authors.affiliation: Zora Biosciences Oy, Biologinkuja 1, FI-02150 Espoo, Finland. kirill.tarasov@outlook.com.
authors.forename: Kirill
authors.initials: K
authors.lastname: Tarasov
authors.forename: Adam
authors.initials: A
authors.lastname: Stefanko
authors.forename: Albert
authors.initials: A
authors.lastname: Casanovas
authors.forename: Michal A
authors.initials: MA
authors.lastname: Surma
authors.forename: Zane
authors.initials: Z
authors.lastname: Berzina
authors.forename: Hans Kristian
authors.initials: HK
authors.lastname: Hannibal-Bach
authors.forename: Kim
authors.initials: K
authors.lastname: Ekroos
authors.forename: Christer S
authors.initials: CS
authors.lastname: Ejsing
mesh_terms: D018929:Cell Culture Techniques; D019295:Computational Biology / Q000379:methods*; D015966:Gene Expression Regulation, Fungal; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D009154:Mutation; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D012984:Software; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D008055:Lipids; D029701:Saccharomyces cerevisiae Proteins