A stable yeast strain efficiently producing cholesterol instead of ergosterol is functional for tryptophan uptake, but not weak organic acid resistance |
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Authors: | Souza Cleiton M Schwabe Tatjana M E Pichler Harald Ploier Birgit Leitner Erich Guan Xue Li Wenk Markus R Riezman Isabelle Riezman Howard |
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Institution: | aDepartment of Biochemistry, University of Geneva, 30 Quai Ernest Ansermet, CH-1211 Geneva, Switzerland;bInstitute of Molecular Biotechnology, Graz University of Technology, Petersgasse 14, A-8010 Graz, Austria;cAustrian Center of Industrial Biotechnology (ACIB), Petersgasse 14, A-8010 Graz, Austria;dInstitute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, A-8010 Graz, Austria;eDepartment of Biochemistry and Department of Biological Sciences, Yong Loo Lin School of Medicine, National University of Singapore, CeLS Building, #04–21, 28 Medical Drive, 117607 Singapore, Singapore |
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Abstract: | Sterols are major lipids in eukaryotes and differ in their specific structure between species. Both cholesterol and ergosterol can form liquid ordered domains in artificial membranes. We reasoned that substituting the main sterol ergosterol by cholesterol in yeast should permit domain formation and discriminate between physical and sterol structure-dependent functions. Using a cholesterol-producing yeast strain, we show that solute transporters for tryptophan and arginine are functional, whereas the export of weak organic acids via Pdr12p, a multi-drug resistance family member, is not. The latter reveals a sterol function that is probably dependent upon a precise sterol structure. We present a series of novel yeast strains with different sterol compositions as valuable tools to characterize sterol function and use them to refine the sterol requirements for Pdr12p. These strains will also be improved hosts for heterologous expression of sterol-dependent proteins and safe sources to obtain pure cholesterol and other sterols. |
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Keywords: | Cholesterol Campesterol Ergosterol DHCR7 DHCR24 Yeast Weak organic acid transport Tryptophan transport Membranes Lipid engineering |
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