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. 2018 Jan 18;8:1020. doi: 10.1038/s41598-018-19360-8

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© The Author(s) 2018

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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The phosphorylated TRBP isoform efficiently reverses PKR’s growth inhibition phenotype in yeast. (A) Yeast growth inhibition assay. Yeast INVSc1 cells were co-transformed with wt PKR/pYES2 and empty vector pYES3CT (PKR alone), wt PKR/pYES2 and K296R PKR/pYES3CT (PKR+K296R), wt PKR/pYES2 and wt TRBP/pYES3CT (PKR+wtTRBP), wt PKR/pYES2 and AAAA TRBP/pYES3CT (PKR+AAAA), or wt PKR/pYES2 and DDDD TRBP/pYES3CT (PKR + DDDD). Ten microliters of transformed yeast cells (OD₆₀₀ = 10, 1, 0.1, 0.01) were spotted on double dropout media (-uracil, - tryptophan) with either glucose (+GLU) or galactose (+GAL) as sole carbon source. Plates were incubated for three days at 30 °C. Transformation of INVSc1 with wt PKR/pYES2 and empty vector pYES3CT served as a control showing growth inhibition on galactose plates, while transformation with wtPKR/pYES2 and K296R PKR/pYES3CT served as a positive control for inhibition of PKR and a reversal of growth inhibition phenotype.