Yeast sphingolipid bypass mutants as indicators of antifungal agents selectively targeting sphingolipid synthesis
Section snippets
Materials and methods
Strains and growth conditions. Yeast strains SJ21R (MATa ura3-52 leu2-3 ade 1), AGD27-61 (MATa/MATα ura3-52 leu2-3,112 lys2-80amber ade1 lcb1::URA3 SLC1-1 ipc1-1), 4R3 (MATa ura3-52 leu2-3,112 ade1 lcb::URA3 SLC1-1), 7R6 (MATa ura3-52 leu2-3,112 ade1 lcb::URA3 SLC1-1), and 7R4 (MATa ura3-52 leu2-3,112 ade1 lcb::URA3 SLC2-1) were obtained from Dr. Bob Dickson (Univ. Kentucky) and were typically maintained in PYED medium (1% yeast extract, 2% Bacto-peptone, 2% glucose, 50 mM sodium succinate, pH
Results and discussion
Yeast sphingolipid bypass mutants are able to grow in the absence of sphingolipid biosynthesis due to their unusual ability to synthesize novel compensatory phospholipids (Fig. 1). The sphingolipid bypass mutants (SLC strains) carry two mutations. One consists of a deletion of the LCB1 gene, which encodes a subunit of serine palmitoyltransferase (SPT), and results in a lack of activity of this normally essential enzyme. The second mutation is a gain of function mutation, SLC1-1, that affects
Acknowledgements
We thank Bob Dickson (Univ. Kentucky) for strains, Hans Achenbach for providing galbonolide A, and Vince Marshall, Joyce Cialdella, Ming-Shang Kuo, and Barclay Shilliday for purifying samples of australifungin and khafrefungin.
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Present address: Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA.