Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Leuven, Belgium.
Center for Microbiology, VIB, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium.
Biochem J. 2021 Jan 29;478(2):357-375. doi: 10.1042/BCJ20200722.
Multiple starvation-induced, high-affinity nutrient transporters in yeast function as receptors for activation of the protein kinase A (PKA) pathway upon re-addition of their substrate. We now show that these transceptors may play more extended roles in nutrient regulation. The Gap1 amino acid, Mep2 ammonium, Pho84 phosphate and Sul1 sulfate transceptors physically interact in vitro and in vivo with the PKA-related Sch9 protein kinase, the yeast homolog of mammalian S6 protein kinase and protein kinase B. Sch9 is a phosphorylation target of TOR and well known to affect nutrient-controlled cellular processes, such as growth rate. Mapping with peptide microarrays suggests specific interaction domains in Gap1 for Sch9 binding. Mutagenesis of the major domain affects the upstart of growth upon the addition of L-citrulline to nitrogen-starved cells to different extents but apparently does not affect in vitro binding. It also does not correlate with the drop in L-citrulline uptake capacity or transceptor activation of the PKA target trehalase by the Gap1 mutant forms. Our results reveal a nutrient transceptor-Sch9-TOR axis in which Sch9 accessibility for phosphorylation by TOR may be affected by nutrient transceptor-Sch9 interaction under conditions of nutrient starvation or other environmental challenges.
酵母中多种因饥饿诱导产生的、高亲和性的营养转运蛋白作为其底物再添加时蛋白激酶 A(PKA)途径激活的受体发挥作用。我们现在表明,这些转运受体可能在营养调节中发挥更广泛的作用。Gap1 氨基酸、Mep2 铵、Pho84 磷酸和 Sul1 硫酸盐转运受体在体外和体内与 PKA 相关的 Sch9 蛋白激酶、哺乳动物 S6 蛋白激酶和蛋白激酶 B 的酵母同源物相互作用。Sch9 是 TOR 的磷酸化靶标,众所周知,它会影响营养控制的细胞过程,例如生长速度。肽微阵列的作图表明 Gap1 中存在用于 Sch9 结合的特定相互作用结构域。主要结构域的突变在添加 L-瓜氨酸到氮饥饿细胞时,以不同的程度影响生长的起始,但显然不影响体外结合。它也与 L-瓜氨酸摄取能力的下降或 Gap1 突变形式的 PKA 靶标海藻糖酶的转运受体激活无关。我们的结果揭示了一个营养转运受体-Sch9-TOR 轴,其中在营养饥饿或其他环境挑战条件下,营养转运受体-Sch9 相互作用可能会影响 TOR 对 Sch9 的磷酸化作用。