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. 2018 Sep 7;7:e35447. doi: 10.7554/eLife.35447

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© 2018, Wang et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 1. — (A) A mot1-301 strain was transformed with CEN LEU2 vectors carrying indicated genes, then selected for transformants on SC-Leu plates. Wild-type MOT1 or SMT3-Q56K made mot1-301 cells grow faster. (B) Transformants from (A) were patched on SC-Leu (-L) then replica plated to SC-Leu-His (-LH), YPSucrose (Suc), SC-Galactose (Gal) plates, or a SC-Leu plate incubated at elevated temperature 38°C (-L 38°C). mot1-301 is His⁺ (Spt^-), Suc⁺ (Bur^-), Gal^-, and Ts^-, whereas SMT3-Q56K reversed all four phenotype. The suppression is dominant because the wild-type genomic copy of SMT3 was present in all the strains. (C) Design of the screen. The starting strain is an ura3 ade2 ade3 triple mutant. ura3 is used for URA3 plasmid selection and 5FOA-sensitivity test. ade2 ade3 double mutant colonies are white, but the wild-type ADE3 on the plasmid complements ade3 and turns the cells red. A mutant (d-) that requires the plasmid for viability will form a uniformly red and 5FOA-sensitive colony.

Figure 1—figure supplement 1. — (A) A mot1-301 strain was transformed with CEN LEU2 vectors carrying indicated genes, then selected for transformants on SC-Leu plates. Wild-type MOT1 or SMT3-Q56K made mot1-301 cells grow faster. (B) Transformants from (A) were patched on SC-Leu (-L) then replica plated to SC-Leu-His (-LH), YPSucrose (Suc), SC-Galactose (Gal) plates, or a SC-Leu plate incubated at elevated temperature 38°C (-L 38°C). mot1-301 is His⁺ (Spt^-), Suc⁺ (Bur^-), Gal^-, and Ts^-, whereas SMT3-Q56K reversed all four phenotype. The suppression is dominant because the wild-type genomic copy of SMT3 was present in all the strains. (C) Design of the screen. The starting strain is an ura3 ade2 ade3 triple mutant. ura3 is used for URA3 plasmid selection and 5FOA-sensitivity test. ade2 ade3 double mutant colonies are white, but the wild-type ADE3 on the plasmid complements ade3 and turns the cells red. A mutant (d-) that requires the plasmid for viability will form a uniformly red and 5FOA-sensitive colony.