A comprehensive protein-protein interactome for yeast PAS kinase 1 reveals direct inhibition of respiration through the phosphorylation of Cbf1.
A comprehensive protein-protein interactome for yeast PAS kinase 1 reveals direct inhibition of respiration through the phosphorylation of Cbf1.
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DOI：
10.1091/mbc.e13-10-0631
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发表时间：
2014-07-15
影响因子：
3.3
通讯作者：
Grose JH
中科院分区：
文献类型：
--
作者：
DeMille D;Bikman BT;Mathis AD;Prince JT;Mackay JT;Sowa SW;Hall TD;Grose JH
PAS激酶是葡萄糖稳态所需的保守的感觉蛋白激酶。酵母PAS激酶1（Psk 1）的相互作用组被确定，揭示了93个结合伴侣。提供了证据表明体内Cbf 1磷酸化并随后抑制呼吸，支持Psk 1在分配葡萄糖以促进细胞生长方面发挥作用。Per-Arnt-Sim（PAS）激酶是酵母、小鼠和人类葡萄糖稳态所需的感觉蛋白激酶，但对其功能的分子机制知之甚少。使用酵母双杂交和共纯化方法，我们确定了酵母PAS激酶1（Psk 1）的蛋白质-蛋白质相互作用组，揭示了93个新的推定蛋白质结合伴侣。几个Psk 1结合伙伴扩大PAS激酶在葡萄糖稳态中的作用，包括参与线粒体代谢的新途径。此外，相互作用组表明PAS激酶在细胞生长（基因/蛋白质表达，复制/细胞分裂，蛋白质修饰和降解），液泡功能和胁迫耐受性中的新作用。在体外激酶研究中，使用这些结合伴侣的25个子集确定Mot 3，Zds 1，Utr 1和Cbf 1作为底物。进一步的证据提供了在T211/T212的Cbf 1的体内磷酸化和随后的呼吸抑制。PAS激酶的这种呼吸作用与PAS激酶缺陷小鼠的高代谢报告一致，确定了可能的分子机制，并巩固了PAS激酶在葡萄糖稳态调节中的进化重要性。
PAS kinase is a conserved sensory protein kinase required for glucose homeostasis. The interactome for yeast PAS kinase 1 (Psk1) is identified, revealing 93 binding partners. Evidence is provided for in vivo phosphorylation of Cbf1 and subsequent inhibition of respiration, supporting a role for Psk1 in partitioning glucose for cell growth. Per-Arnt-Sim (PAS) kinase is a sensory protein kinase required for glucose homeostasis in yeast, mice, and humans, yet little is known about the molecular mechanisms of its function. Using both yeast two-hybrid and copurification approaches, we identified the protein–protein interactome for yeast PAS kinase 1 (Psk1), revealing 93 novel putative protein binding partners. Several of the Psk1 binding partners expand the role of PAS kinase in glucose homeostasis, including new pathways involved in mitochondrial metabolism. In addition, the interactome suggests novel roles for PAS kinase in cell growth (gene/protein expression, replication/cell division, and protein modification and degradation), vacuole function, and stress tolerance. In vitro kinase studies using a subset of 25 of these binding partners identified Mot3, Zds1, Utr1, and Cbf1 as substrates. Further evidence is provided for the in vivo phosphorylation of Cbf1 at T211/T212 and for the subsequent inhibition of respiration. This respiratory role of PAS kinase is consistent with the reported hypermetabolism of PAS kinase–deficient mice, identifying a possible molecular mechanism and solidifying the evolutionary importance of PAS kinase in the regulation of glucose homeostasis.