García Patricia, Encinar Del Dedo Javier, Ayté José, Hidalgo Elena
From the Oxidative Stress and Cell Cycle Group, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, 08003 Barcelona, Spain.
From the Oxidative Stress and Cell Cycle Group, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, 08003 Barcelona, Spain
J Biol Chem. 2016 Jan 8;291(2):790-9. doi: 10.1074/jbc.M115.696658. Epub 2015 Nov 13.
In response to environmental cues, the mitogen-activated protein kinase Sty1-driven signaling cascade activates hundreds of genes to induce a robust anti-stress cellular response in fission yeast. Thus, upon stress imposition Sty1 transiently accumulates in the nucleus where it up-regulates transcription through the Atf1 transcription factor. Several regulators of transcription and translation have been identified as important to mount an integral response to oxidative stress, such as the Spt-Ada-Gcn5-acetyl transferase or Elongator complexes, respectively. With the aim of identifying new regulators of this massive gene expression program, we have used a GFP-based protein reporter and screened a fission yeast deletion collection using flow cytometry. We find that the levels of catalase fused to GFP, both before and after a threat of peroxides, are altered in hundreds of strains lacking components of chromatin modifiers, transcription complexes, and modulators of translation. Thus, the transcription elongation complex Paf1, the histone methylase Set1-COMPASS, and the translation-related Trm112 dimers are all involved in full expression of Ctt1-GFP and in wild-type tolerance to peroxides.
作为对环境信号的响应,丝裂原活化蛋白激酶Sty1驱动的信号级联反应激活数百个基因,以在裂殖酵母中诱导强大的抗应激细胞反应。因此,在施加应激时,Sty1会短暂积累在细胞核中,通过Atf1转录因子上调转录。转录和翻译的几种调节因子已被确定为对氧化应激产生整体反应很重要,例如分别为Spt-Ada-Gcn5-乙酰转移酶或延伸因子复合物。为了鉴定这个大规模基因表达程序的新调节因子,我们使用了基于绿色荧光蛋白(GFP)的蛋白质报告基因,并通过流式细胞术筛选了裂殖酵母缺失文库。我们发现,在缺乏染色质修饰剂、转录复合物和翻译调节因子成分的数百个菌株中,与GFP融合的过氧化氢酶水平在受到过氧化物威胁之前和之后都会发生改变。因此,转录延伸复合物Paf1、组蛋白甲基转移酶Set1-COMPASS和与翻译相关的Trm112二聚体都参与Ctt1-GFP的完全表达以及对过氧化物的野生型耐受性。