Amino acid residues involved in ligand preference of the Snf3 transporter-like sensor in Saccharomyces cerevisiae

J. Dietvorst, Kaisa Karhumaa, Morten Kielland-Brandt, Anders Bøving Brandt

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Snf3 is a plasma membrane protein in Saccharomyces cerevisiae cerevisiae able to sense the presence of glucose. Although the Snf3 protein does not transport sugars, it shares sequence similarity with various glucose transporters from other organisms. we investigated the sugar specificity/preferences of Snf3. The ability of cells to sense sugars in vivo was monitored by following the degradation of the Mth1 protein, :ill earl., event ill the signal pathway. Our study reveals that Snf3. ill addition to glucose. also senses fructose and mannose, as well as the glucose analogues 2-deoxyglucose, 3-O-methylglucoside and 6-deoxyglucose. The signalling proficiency of a non-phosphorylatable analogue strongly supports the notion that sensing through Snf3 does not require sugar phosphorylation. Sequence comparisons of Snf3 to glucose transporters indicated amino acid residues possibly involved in sensing of sugars other than glucose. By site-specific mutagenesis of the structural gene, roles of specific residues in Snf3 could he established. Change of isoleucine-374 to valine ill transmembrane segment 7 of Snf3 partially abolished sensing of fructose mannose. while mutagenesis causing it change of phenylalanine-462 (4) tyrosine ill transmembrane segment 10 of Snf3 abolished sensing of fructose. Neither of these amino :kill changes affected the ability of Snf3 to sense glucose, nor did they permit Snf3 to sense galactose. These data indicate it similarity between it ligand binding site of the sensor Snf3 and binding sites used for facilitated hexose transport in the GLUT proteins.
    Original languageEnglish
    JournalYeast
    Volume27
    Issue number3
    Pages (from-to)131-138
    ISSN0749-503X
    DOIs
    Publication statusPublished - 2010

    Keywords

    • glucose sensing
    • mannose sensing
    • fructose sensing
    • yeast
    • Snf3 transporter-like sensor

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