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. 2017 Oct 23;7:13816. doi: 10.1038/s41598-017-14124-2

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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FTY720 affects the activity of various permeases. (A) Strains 27038a (rsp5(npi1) ura3), CJ005 (gap1Δ rsp5(npi1) ura3), and 35288a (gap1Δ can1Δ rsp5(npi1) ura3) were transformed with plasmid pFL38 (URA3). Cells were treated or not with FTY720 (10 μM) or rapamycin (200 ng/ml) for 1 h. The initial uptake rate for 75 μM [¹⁴C]-labeled citrulline (reflecting Gap1 activity), 10 μM [¹⁴C]-labeled arginine (reflecting Can1 activity), 10 μM [¹⁴C]-labeled lysine (reflecting Lyp1 activity), or 5 μM [¹⁴C]-labeled uracil (reflecting Fur4 activity) was then measured. Error bars correspond to SD, n = 3–5 (unpaired t-test, *P < 0.05, **P < 0.01, ***P < 0.001). (B) Strain EK008 (gap1Δ ura3) transformed with plasmid pCJ038 (YCpGAL-GAP1^K9R,K16R-GFP) was grown on galactose-proline medium. Glucose was added for 90 min to inhibit Gap1 synthesis. Cells were treated for 0, 10, 30, 60, or 120 min with FTY720 (10 μM final concentration) or DMSO alone (control). The initial uptake rate of [¹⁴C]-labeled citrulline (75 μM), reflecting Gap1 activity, was then measured. Error bars correspond to SD, n = 2. (C) Strain EK008 (gap1Δ ura3) transformed with pCJ038 (YCpGAL-GAP1^K9R,K16R-GFP) was grown on galactose-proline medium. Glucose was added for 90 min to repress Gap1 synthesis. Cells were treated for 60 min with FTY720 or the solvent DMSO alone (control) and the uptake of [¹⁴C]-citrulline added at various concentrations was then measured. Error bars correspond to SD, n = 2.