Guarini Nadia, Saliba Elie, André Bruno
Molecular Physiology of the Cell, Université Libre de Bruxelles (ULB), Biopark, Gosselies, Belgium.
PLoS Genet. 2024 Jan 16;20(1):e1011121. doi: 10.1371/journal.pgen.1011121. eCollection 2024 Jan.
Plasma membrane (PM) H+-ATPases of the P-type family are highly conserved in yeast, other fungi, and plants. Their main role is to establish an H+ gradient driving active transport of small ions and metabolites across the PM and providing the main component of the PM potential. Furthermore, in both yeast and plant cells, conditions have been described under which active H+-ATPases promote activation of TORC1, the rapamycin-sensitive kinase complex controlling cell growth. Fungal and plant PM H+-ATPases are self-inhibited by their respective cytosolic carboxyterminal tails unless this domain is phosphorylated at specific residues. In the yeast H+-ATPase Pma1, neutralization of this autoinhibitory domain depends mostly on phosphorylation of the adjacent Ser911 and Thr912 residues, but the kinase(s) and phosphatase(s) controlling this tandem phosphorylation remain unknown. In this study, we show that S911-T912 phosphorylation in Pma1 is mediated by the largely redundant Ptk1 and Ptk2 kinase paralogs. Dephosphorylation of S911-T912, as occurs under glucose starvation, is dependent on the Glc7 PP1 phosphatase. Furthermore, proper S911-T912 phosphorylation in Pma1 is required for optimal TORC1 activation upon H+ influx coupled amino-acid uptake. We finally show that TORC1 controls S911-T912 phosphorylation in a manner suggesting that activated TORC1 promotes feedback inhibition of Pma1. Our results shed important new light on phosphoregulation of the yeast Pma1 H+-ATPase and on its interconnections with TORC1.
P型家族的质膜(PM)H⁺-ATP酶在酵母、其他真菌和植物中高度保守。它们的主要作用是建立一个H⁺梯度,驱动小离子和代谢物跨质膜的主动运输,并提供质膜电位的主要成分。此外,在酵母和植物细胞中,都描述了活性H⁺-ATP酶促进TORC1激活的条件,TORC1是一种对雷帕霉素敏感的激酶复合物,控制细胞生长。真菌和植物的质膜H⁺-ATP酶被其各自的胞质羧基末端尾巴自我抑制,除非该结构域在特定残基处被磷酸化。在酵母H⁺-ATP酶Pma1中,这种自抑制结构域的中和主要取决于相邻的Ser911和Thr912残基的磷酸化,但控制这种串联磷酸化的激酶和磷酸酶仍然未知。在本研究中,我们表明Pma1中S911-T912的磷酸化是由高度冗余的Ptk1和Ptk2激酶旁系同源物介导的。如在葡萄糖饥饿条件下发生的S911-T912去磷酸化依赖于Glc7 PP1磷酸酶。此外,Pma1中适当的S911-T912磷酸化是H⁺流入偶联氨基酸摄取后TORC1最佳激活所必需的。我们最终表明,TORC1以一种表明激活的TORC1促进Pma1反馈抑制的方式控制S911-T912的磷酸化。我们的结果为酵母Pma1 H⁺-ATP酶的磷酸调节及其与TORC1的相互关系提供了重要的新见解。