酵母氮同化过程中通用氨基酸控制和雷帕霉素靶蛋白(TOR)调节途径的整合。

Integration of general amino acid control and target of rapamycin (TOR) regulatory pathways in nitrogen assimilation in yeast.

机构信息

Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.

出版信息

J Biol Chem. 2010 May 28;285(22):16893-911. doi: 10.1074/jbc.M110.121947. Epub 2010 Mar 16.

Abstract

Two important nutrient-sensing and regulatory pathways, the general amino acid control (GAAC) and the target of rapamycin (TOR), participate in the control of yeast growth and metabolism during changes in nutrient availability. Amino acid starvation activates the GAAC through Gcn2p phosphorylation of translation factor eIF2 and preferential translation of GCN4, a transcription activator. TOR senses nitrogen availability and regulates transcription factors such as Gln3p. We used microarray analyses to address the integration of the GAAC and TOR pathways in directing the yeast transcriptome during amino acid starvation and rapamycin treatment. We found that GAAC is a major effector of the TOR pathway, with Gcn4p and Gln3p each inducing a similar number of genes during rapamycin treatment. Although Gcn4p activates a common core of 57 genes, the GAAC directs significant variations in the transcriptome during different stresses. In addition to inducing amino acid biosynthetic genes, Gcn4p in conjunction with Gln3p activates genes required for the assimilation of secondary nitrogen sources such as gamma-aminobutyric acid (GABA). Gcn2p activation upon shifting to secondary nitrogen sources is suggested to occur by means of a dual mechanism. First, Gcn2p is induced by the release of TOR repression through a mechanism involving Sit4p protein phosphatase. Second, this eIF2 kinase is activated by select uncharged tRNAs, which were shown to accumulate during the shift to the GABA medium. This study highlights the mechanisms by which the GAAC and TOR pathways are integrated to recognize changing nitrogen availability and direct the transcriptome for optimal growth adaptation.

摘要

两种重要的营养感应和调节途径,即一般氨基酸控制 (GAAC) 和雷帕霉素靶蛋白 (TOR),参与了酵母在营养供应变化时的生长和代谢控制。氨基酸饥饿通过 Gcn2p 磷酸化翻译因子 eIF2 和 GCN4 的优先翻译激活 GAAC,GCN4 是一种转录激活因子。TOR 感应氮的可用性,并调节 Gln3p 等转录因子。我们使用微阵列分析来解决 GAAC 和 TOR 途径在氨基酸饥饿和雷帕霉素处理期间指导酵母转录组的整合。我们发现 GAAC 是 TOR 途径的主要效应因子,在雷帕霉素处理期间,Gcn4p 和 Gln3p 各自诱导相似数量的基因。尽管 Gcn4p 激活了 57 个基因的共同核心,但 GAAC 在不同应激条件下指导转录组的显著变化。除了诱导氨基酸生物合成基因外,Gcn4p 与 Gln3p 一起激活了用于同化次要氮源(如γ-氨基丁酸 (GABA))的基因。当转向次要氮源时,Gcn2p 的激活被认为是通过双重机制发生的。首先,通过 Sit4p 蛋白磷酸酶涉及的机制,通过释放 TOR 抑制来诱导 Gcn2p。其次,这种 eIF2 激酶被选择的无电荷 tRNA 激活,这些 tRNA 在向 GABA 培养基转移时积累。这项研究强调了 GAAC 和 TOR 途径如何整合以识别不断变化的氮可用性并指导转录组以实现最佳生长适应的机制。

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