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verse species of fatty acyl chains. Remodeling of fatty acyl chains is supposed to be vital for the maintenance of membrane structure and function. However, the mechanism and physiological roles of phospholipid remodeling are not fully clarified. To elucidate the mechanism of phospholipid remodeling, it is effective to establish a system in which exogenous phospholipids or phospholipid analogs are incorporated into the cells and to monitor their metabolism in the cells. Phosphatidylethanolamine (PE) is one of the major and essential nonbilayer phospholipids and comprises ~20% of total phospholipids in eukaryotic cells (Daum et al., 1999; Storey et al., 2001). We have previously constructed a Saccharomyces cerevisiae strain, TKY12Ga, that is defective in phosphatidylserine (PS) decarboxylation by deletion of PSD1 and PSD2, and its PE synthesis can be controlled by the carbon source in the media through replacement of the promoter of ECT1, encoding CTP: phosphoethanolamine cytidylyltransferase, a key enzyme in PE synthesis through the Kennedy pathway, with the galactose-inducible and glucose-repressible GAL1 promoter (MinSeok et al., 1996; Deng, L., unpubl.). In galactose-containing minimal medium (SG medium), TKY12Ga grew in the presence of ethanolamine. In contrast, its growth was inhibited in the glucose-containing minimal medium (SD medium), even in the presence of ethanolamine. However, when didecanoyl PE (diC10PE) was added to the SD medium, TKY12Ga grew despite the inability to synthesize PE. Since the length of acyl chains in PE in yeast is mainly C14 to C18, it was presumed that the PE with short acyl chains was remodeled to those containing acyl chains of normal length to be a component in the biological membrane. Here, to obtain information on the utilization of exogenous PE in yeast, we isolated mutants defective in utilization of exogenous PE to support growth when the synthesis of PE was suppressed, and analyzed one of them. We mutagenized TKY12Ga with ethyl methanesulfonate (EMS) and isolated thirty-three mutants defective in growth on SD medium containing diC10PE from approximately 63,000 colonies. Among those, M25, M30, and M33 exhibited clear and stable phenotypes. These mutants did not grow in SD medium containing diC10PE, while they grew normally in SG medium containing ethanolamine at 30°C. In addition, M25 and M30 had the phenotype of temperature sensitivity in SG medium at 37°C (Fig. 2B–D, data not shown). However, the phenotypes of M30 and M33 were caused by multiple mutations, while that of M25 was by a single mutation (data not shown, see below). Therefore, M25 was chosen for further analysis. Isolation and characterization of a mutant defective in utilization of exogenous phosphatidylethanolamine in Saccharomyces cerevisiae