Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Department of Biology and Genetics Institute, University of Florida, Gainesville, Florida, USA.
Aging Cell. 2023 Apr;22(4):e13795. doi: 10.1111/acel.13795. Epub 2023 Feb 16.
CCR4-NOT is a versatile eukaryotic protein complex that controls multiple steps in gene expression regulation from synthesis to decay. In yeast, CCR4-NOT has been implicated in stress response regulation, though this function in other organisms remains unclear. In a genome-wide RNAi screen, we identified a subunit of the CCR4-NOT complex, ccf-1, as a requirement for the C. elegans transcriptional response to cadmium and acrylamide stress. Using whole-transcriptome RNA sequencing, we show that the knockdown of ccf-1 attenuates the activation of a broad range of stress-protective genes in response to cadmium and acrylamide, including those encoding heat shock proteins and xenobiotic detoxification. Consistently, survival assays show that the knockdown of ccf-1 decreases C. elegans stress resistance and normal lifespan. A yeast 2-hybrid screen using a CCF-1 bait identified the homeobox transcription factor PAL-1 as a physical interactor. Knockdown of pal-1 inhibits the activation of ccf-1 dependent stress genes and reduces C. elegans stress resistance. Gene expression analysis reveals that knockdown of ccf-1 and pal-1 attenuates the activation of elt-2 and elt-3 under stress that encode master transcriptional co-regulators of stress response in the C. elegans, and that overexpression of ELT-2 can suppress ccf-1's requirement for gene transcription in a stress-dependent manner. Our findings reveal a new role for CCR4-NOT in the environmental stress response and define its role in stress resistance and longevity in C. elegans.
CCR4-NOT 是一种多功能的真核蛋白复合物,可控制从合成到降解的多个基因表达调控步骤。在酵母中,CCR4-NOT 已被牵连到应激反应调节中,尽管在其他生物中其功能尚不清楚。在全基因组 RNAi 筛选中,我们鉴定了 CCR4-NOT 复合物的一个亚基 ccf-1,作为线虫对镉和丙烯酰胺应激转录反应的必需条件。通过全转录组 RNA 测序,我们发现 ccf-1 的敲低会减弱对镉和丙烯酰胺的广泛应激保护基因的激活,包括编码热休克蛋白和外来物解毒的基因。一致地,生存测定表明 ccf-1 的敲低会降低线虫的应激抗性和正常寿命。使用 CCF-1 诱饵的酵母 2 杂交筛选鉴定了同源盒转录因子 PAL-1 作为物理相互作用者。pal-1 的敲低抑制 ccf-1 依赖的应激基因的激活,并降低线虫的应激抗性。基因表达分析表明,ccf-1 和 pal-1 的敲低会减弱在应激条件下 elt-2 和 elt-3 的激活,编码线虫应激反应的主转录共调节因子;并且 ELT-2 的过表达可以以应激依赖的方式抑制 ccf-1 在基因转录中的需求。我们的发现揭示了 CCR4-NOT 在环境应激反应中的新作用,并定义了其在线虫应激抗性和寿命中的作用。