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Inge Holsbeeks

Publications and source records attributed to Inge Holsbeeks.

2 recordsLinked to original sources

The eukaryotic plasma membrane as a nutrient-sensing device.

In eukaryotic cells, G-protein-coupled receptors (GPCRs), non-transporting nutrient carrier homologues and active nutrient carriers have been recently shown to function as sensors that directly monitor the level of nutrients in the extracellular environment. The plasma membrane is not only the cellular boundary at which signalling molecules that govern metabolism and proliferation are detected, but also the boundary across which nutrients that sustain the generation of energy and building blocks are transported. Nutrient sensors combine these functions in various ways. Classical receptor proteins detect the presence of nutrients, carriers combine the functions of nutrient transporters and receptors, and carrier homologues have lost their transport capacity and become pure receptors. The activation of signal transduction pathways by nutrients adds a new layer to the regulatory network that controls metabolism and proliferation. Nutrient sensors highlight the importance of both nutrients as signalling molecules and nutrient carriers as receptors for signalling pathways.

Biological Transport↗

The Gap1 general amino acid permease acts as an amino acid sensor for activation of protein kinase A targets in the yeast Saccharomyces cerevisiae.

Addition of a nitrogen source to yeast (Saccharomyces cerevisiae) cells starved for nitrogen on a glucose-containing medium triggers activation of protein kinase A (PKA) targets through a pathway that requires for sustained activation both a fermentable carbon source and a complete growth medium (fermentable growth medium induced or FGM pathway). Trehalase is activated, trehalose and glycogen content as well as heat resistance drop rapidly, STRE-controlled genes are repressed, and ribosomal protein genes are induced. We show that the rapid effect of amino acids on these targets specifically requires the general amino acid permease Gap1. In the gap1Delta strain, transport of high concentrations of l-citrulline occurs at a high rate but without activation of trehalase. Metabolism of the amino acids is not required. Point mutants in Gap1 with reduced or deficient transport also showed reduced or deficient signalling. However, two mutations, S391A and S397A, were identified with a differential effect on transport and signalling for l-glutamate and l-citrulline. Specific truncations of the C-terminus of Gap1 (e.g. last 14 or 26 amino acids) did not reduce transport activity but caused the same phenotype as in strains with constitutively high PKA activity also during growth with ammonium as sole nitrogen source. The overactive PKA phenotype was abolished by mutations in the Tpk1 or Tpk2 catalytic subunits. We conclude that Gap1 acts as an amino acid sensor for rapid activation of the FGM signalling pathway which controls the PKA targets, that transport through Gap1 is connected to signalling and that specific truncations of the C-terminus result in permanently activating Gap1 alleles.

Amino Acid Sequence↗