Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa 277-8562, Japan.
Division of Biotechnology, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-0851, Japan.
Int J Mol Sci. 2021 May 28;22(11):5798. doi: 10.3390/ijms22115798.
The Pv11, an insect cell line established from the midge , is capable of extreme hypometabolic desiccation tolerance, so-called anhydrobiosis. We previously discovered that heat shock factor 1 (HSF1) contributes to the acquisition of desiccation tolerance by Pv11 cells, but the mechanistic details have yet to be elucidated. Here, by analyzing the gene expression profiles of newly established HSF1-knockout and -rescue cell lines, we show that HSF1 has a genome-wide effect on gene regulation in Pv11. The HSF1-knockout cells exhibit a reduced desiccation survival rate, but this is completely restored in HSF1-rescue cells. By comparing mRNA profiles of the two cell lines, we reveal that HSF1 induces anhydrobiosis-related genes, especially genes encoding late embryogenesis abundant proteins and thioredoxins, but represses a group of genes involved in basal cellular processes, thus promoting an extreme hypometabolism state in the cell. In addition, HSF1 binding motifs are enriched in the promoters of anhydrobiosis-related genes and we demonstrate binding of HSF1 to these promoters by ChIP-qPCR. Thus, HSF1 directly regulates the transcription of anhydrobiosis-related genes and consequently plays a pivotal role in the induction of anhydrobiotic ability in Pv11 cells.
从摇蚊中建立的昆虫细胞系 Pv11 能够实现极端的低代谢干燥耐受性,即所谓的脱水休眠。我们之前发现热休克因子 1(HSF1)有助于 Pv11 细胞获得干燥耐受性,但具体的机制细节尚未阐明。在这里,通过分析新建立的 HSF1 敲除和挽救细胞系的基因表达谱,我们表明 HSF1 对 Pv11 中的基因调控具有全基因组的影响。HSF1 敲除细胞表现出降低的干燥存活率,但在 HSF1 挽救细胞中完全恢复。通过比较这两种细胞系的 mRNA 谱,我们揭示了 HSF1 诱导了与脱水休眠相关的基因,特别是编码晚期胚胎丰富蛋白和硫氧还蛋白的基因,但抑制了一组涉及基础细胞过程的基因,从而促进了细胞的极度低代谢状态。此外,HSF1 结合基序在与脱水休眠相关的基因启动子中富集,我们通过 ChIP-qPCR 证明了 HSF1 与这些启动子的结合。因此,HSF1 直接调节与脱水休眠相关基因的转录,从而在 Pv11 细胞中诱导脱水休眠能力中发挥关键作用。