Human wild-type and pathogenic TPI variants can substitute for yeast TPI1.

Abstract

<div><p> <b>A)</b> MR100 yeast cells (<i>Δtpi1</i>) were transformed with the various p416GPD-based expression plasmids encoding wild-type human TPI as well as the pathogenic variants Met1_AAG, Cys41Tyr, Glu104Asp, Gly122Arg, Ile170Val or Phe240Leu, respectively, and plated on minimal SC ^-leu-ura medium supplemented with 3% ethanol/0.1% glucose.</p> <p>Afterwards, single yeast clones were selected and grown as represented by the schemes on SC ^-leu-ura medium plates supplemented either with 2% glucose or with 3% ethanol/0.1% glucose at 30°C. <b>B)</b> MR100 <i>Δtpi1</i> yeast cells expressing wild-type TPI or the different pathogenic TPI variants were grown until logarithmic phase.</p> <p>Then, the same cell number of each culture was spotted as 5-fold serial dilutions onto glucose media or onto glucose media supplemented with different concentrations of lithium chloride. </p> <p>Plates were incubated for 3 days at 30°C and growth of the different yeast strains was analyzed.</p></div