MPM motifs of the yeast SKT protein Trk1 can assemble to form a functional K+-translocation system.

Katsiaryna Shamayeva, Karin Spurna, Natalia Kulik, Deepika Kale, Oksana Munko, Pavel Spurny, Vasilina Zayats, Jost Ludwig

Biochimica et biophysica acta. Biomembranes 2021 Feb 01

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The yeast Trk1 polypeptide, like other members of the Superfamily of K Transporters (SKT proteins) consists of four Membrane-Pore-Membrane motifs (MPMs A-D) each of which is homologous to a single K-channel subunit. SKT proteins are thought to have evolved from ancestral K-channels via two gene duplications and thus single MPMs might be able to assemble when located on different polypeptides. To test this hypothesis experimentally we generated a set of partial gene deletions to create alleles encoding one, two, or three MPMs, and analysed the cellular localisation and interactions of these Trk1 fragments using GFP tags and Bimolecular Fluorescence Complementation (BiFC). The function of these partial Trk1 proteins either alone or in combinations was assessed by expressing the encoding genes in a K+-uptake deficient strain lacking also the K-channel Tok1 (trk1,trk2,tok1Δ) and (i) analysing their ability to promote growth in low [K+] media and (ii) by ion flux measurements using "microelectrode based ion flux estimation" (MIFE). We found that proteins containing only one or two MPM motifs can interact with each other and assemble with a polypeptide consisting of the rest of the Trk system to form a functional K+-translocation system. Copyright © 2020 Elsevier B.V. All rights reserved.

Citation

Katsiaryna Shamayeva, Karin Spurna, Natalia Kulik, Deepika Kale, Oksana Munko, Pavel Spurny, Vasilina Zayats, Jost Ludwig. MPM motifs of the yeast SKT protein Trk1 can assemble to form a functional K+-translocation system. Biochimica et biophysica acta. Biomembranes. 2021 Feb 01;1863(2):183513