Alteration of the lipid composition of biological membranes interferes with their function and can cause tissue damage by triggering apoptosis. Upon lipid bilayer stress, the endoplasmic reticulum mounts a stress response that is similar to the unfolded protein response. However, only a few genes are known to regulate lipid bilayer stress. Here, we performed a suppressor screen that combined the auxin-inducible degradation (AID) system with conventional RNAi in C. elegans to identify members of the lipid bilayer stress response. AID-mediated knockdown of the mediator MDT-15, a protein required for the upregulation of fatty acid desaturases, caused activation of a lipid bilayer stress sensitive reporters. We screened through almost all C. elegans kinases and transcription factors using RNAi by feeding. We report the identification of 8 genes that have not been implicated previously with lipid bilayer stress before in C. elegans. These suppressor genes include skn-1/NRF1,2,3 and let-607/CREB3. Our candidate suppressor genes suggest a network of transcription factors and the integration of multiple tissues for a centralized lipotoxicity response in the intestine. Additionally, we propose and demonstrate the proof-of-concept for combining AID and RNAi as a new screening strategy.

中文翻译：

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生长素诱导的降解和RNAi筛选相结合，确定了秀丽隐杆线虫脂质双层应激感测中涉及的新基因

生物膜脂质组成的改变会干扰其功能，并可能通过触发细胞凋亡而引起组织损伤。在脂质双层压力作用下，内质网产生的压力反应与未折叠的蛋白质反应相似。然而，已知仅有少数基因可调节脂质双层压力。在这里，我们进行了抑制剂筛选，将线虫中的生长素诱导降解（AID）系统与常规RNAi相结合，以鉴定脂质双层应激反应的成员。AID介导的介质MDT-15（脂肪酸去饱和酶上调所必需的一种蛋白）的敲低导致脂质双层应激敏感报道分子的激活。我们筛选了几乎所有的秀丽隐杆线虫进食时使用RNAi合成激酶和转录因子。我们报告鉴定的线虫中以前没有涉及脂质双层应激的8个基因的鉴定。这些抑制基因包括skn-1 / NRF1,2,3和let-607 / CREB3。我们的候选抑制基因表明，转录因子网络和多个组织的整合可在肠道内实现集中的脂毒性反应。此外，我们提出并证明了将AID和RNAi结合起来作为新的筛选策略的概念证明。