Kingsbury Joanne M, Cardenas Maria E
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710.
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina 27710
G3 (Bethesda). 2016 Jan 6;6(3):641-52. doi: 10.1534/g3.115.023911.
The Target of Rapamycin Complex I (TORC1) orchestrates global reprogramming of transcriptional programs in response to myriad environmental conditions, yet, despite the commonality of the TORC1 complex components, different TORC1-inhibitory conditions do not elicit a uniform transcriptional response. In Saccharomyces cerevisiae, TORC1 regulates the expression of nitrogen catabolite repressed (NCR) genes by controlling the nuclear translocation of the NCR transactivator Gln3. Moreover, Golgi-to-endosome trafficking was shown to be required for nuclear translocation of Gln3 upon a shift from rich medium to the poor nitrogen source proline, but not upon rapamycin treatment. Here, we employed microarray profiling to survey the full impact of the vesicular trafficking system on yeast TORC1-orchestrated transcriptional programs. In addition to the NCR genes, we found that ribosomal protein, ribosome biogenesis, phosphate-responsive, and sulfur-containing amino acid metabolism genes are perturbed by disruption of Golgi-to-endosome trafficking following a nutritional shift from rich to poor nitrogen source medium, but not upon rapamycin treatment. Similar to Gln3, defects in Golgi-to-endosome trafficking significantly delayed cytoplasmic-nuclear translocation of Sfp1, but did not detectably affect the cytoplasmic-nuclear or nuclear-cytoplasmic translocation of Met4, which are the transactivators of these genes. Thus, Golgi-to-endosome trafficking defects perturb TORC1 transcriptional programs via multiple mechanisms. Our findings further delineate the downstream transcriptional responses of TORC1 inhibition by rapamycin compared with a nitrogen quality downshift. Given the conservation of both TORC1 and endomembrane networks throughout eukaryotes, our findings may also have implications for TORC1-mediated responses to nutritional cues in mammals and other eukaryotes.
雷帕霉素复合体I(TORC1)可根据多种环境条件协调转录程序的全局重编程,然而,尽管TORC1复合体的组成成分具有共性,但不同的TORC1抑制条件并不会引发统一的转录反应。在酿酒酵母中,TORC1通过控制氮分解代谢物阻遏(NCR)转录激活因子Gln3的核转位来调节NCR基因的表达。此外,研究表明,从丰富培养基转变为贫氮源脯氨酸时,高尔基体到内体的转运对于Gln3的核转位是必需的,但雷帕霉素处理时并非如此。在这里,我们利用微阵列分析来全面研究囊泡运输系统对酵母TORC1协调的转录程序的影响。除了NCR基因外,我们发现,从丰富氮源培养基转变为贫氮源培养基后,高尔基体到内体的运输受到破坏会扰乱核糖体蛋白、核糖体生物发生、磷酸盐响应和含硫氨基酸代谢基因,但雷帕霉素处理时不会。与Gln3类似,高尔基体到内体运输的缺陷显著延迟了Sfp1的细胞质-核转位,但未检测到对这些基因的转录激活因子Met4的细胞质-核或核-细胞质转位有影响。因此,高尔基体到内体的运输缺陷通过多种机制扰乱TORC1转录程序。我们的研究结果进一步描绘了与氮质量下降相比,雷帕霉素抑制TORC1的下游转录反应。鉴于TORC1和内膜网络在整个真核生物中的保守性,我们的研究结果可能也对哺乳动物和其他真核生物中TORC1介导的营养信号反应有影响。