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. 2015 Feb 10;6:47. doi: 10.3389/fpls.2015.00047

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Copyright © 2015 Safiarian, Pertl-Obermeyer, Lughofer, Hude, Bertl and Obermeyer.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Functional complementation of yeast mutants with modified LilKT1 proteins. (A) Single (LilKT1_K840D) and double mutations (LilKT1_G797D/K840D) of the di-acidic motifs in the C-terminus of LilKT1. (B) Expression of LilKT1 with a total deletion of the C-terminus (ΔCtermLilKT1) and expression of chimeric protein with the N-terminus of LilKT1 and the C-terminus of AKT1 (chimLilKT1) did partially complement the growth deficiency of PLY240 in low K⁺ media. (C) Detection of the C-terminal deleted and chimeric LilKT1 in the cytosolic (CF) and microsomal (MF) fraction of yeast cells (PLY240) using an anti-HA-tag antibody (1:10,000) and a goat anti-mouse IgG AP-conjugated antibody (1:50,000). Predicted molecular weight s for ΔCtermLilKT1 and chimLilKT1 are 47.6 and 106.8 kDa, respectively. Channels were cloned into pGREG535. (D) Fluorescence localization of GFP::yLilKT1/AKT1 chimera in yeast protoplasts (left) and cells (right). Bar = 5 μm.