Targeting of malate synthase 1 to the peroxisomes of Saccharomyces cerevisiae cells depends on growth on oleic acid medium.
The eukaryotic glyoxylate cycle has been previously hypothesized to occur in the peroxisomal compartment, which in the yeast Saccharomyces cerevisiae additionally represents the sole site for fatty acid beta-oxidation. The subcellular location of the key glyoxylate-cycle enzyme malate synthase 1 (Mls1p), an SKL-terminated protein, was examined in yeast cells grown on different carbon sources. Immunoelectron microscopy in combination with cell fractionation showed that Mls1p was abundant in the peroxisomes of cells grown on oleic acid, whereas in ethanol-grown cells Mls1p was primarily cytosolic. This was reinforced using a green fluorescent protein (GFP)-Mls1p reporter, which entered peroxisomes solely in cells grown under oleic acid-medium conditions. Although growth of cells devoid of Mls1p on ethanol or acetate could be fully restored using a cytosolic Mls1p devoid of SKL, this construct could only partially alleviate the requirement for native Mls1p in cells grown on oleic acid. The combined results indicated that Mls1p remained in the cytosol of cells grown on ethanol, and that targeting of Mls1p to the peroxisomes was advantageous to cells grown on oleic acid as a sole carbon source.
Kunze
Markus
M
Institut für Biochemie und Molekulare Zellbiologie der Universität Wien and Ludwig Boltzmann-Forschungsstelle für Biochemie, Vienna Biocenter, Austria.
Kragler
Friedrich
F
Binder
Maximilian
M
Hartig
Andreas
A
Gurvitz
Aner
A
D002455:Cell Division; D003470:Culture Media / Q000737:chemistry; D000431:Ethanol / Q000378:metabolism; D006038:Glyoxylates / Q000378:metabolism; D008292:Malate Synthase / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D019301:Oleic Acid / Q000378:metabolism*; D020675:Peroxisomes / Q000378:metabolism*; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism*
D003470:Culture Media; D006038:Glyoxylates; D019301:Oleic Acid; D000431:Ethanol; D008292:Malate Synthase; C031150:glyoxylic acid