The zoonotic human pathogen Campylobacter jejuni is known for its ability to induce DNA-damage and cell death pathology in humans. The molecular mechanism behind this phenomenon involves nuclear translocation by Cas9, a nuclease in C. jejuni (CjeCas9) that is the molecular marker of the Type II CRISPR-Cas system. However, it is unknown via which cellular pathways CjeCas9 drives human intestinal epithelial cells into cell death. Here, we show that CjeCas9 released by C. jejuni during the infection of Caco-2 human intestinal epithelial cells directly modulates Caco-2 transcriptomes during the first four hours of infection. Specifically, our results reveal that CjeCas9 activates DNA damage (p53, ATM (Ataxia Telangiectasia Mutated Protein)), pro-inflammatory (NF-κB (Nuclear factor-κB)) signaling and cell death pathways, driving Caco-2 cells infected by wild-type C. jejuni, but not when infected by a cas9 deletion mutant, towards programmed cell death. This work corroborates our previous finding that CjeCas9 is cytotoxic and highlights on a RNA level the basal cellular pathways that are modulated.

中文翻译：

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空肠弯曲杆菌 Cas9 调节 Caco-2 肠上皮细胞的转录组

人畜共患病病原体空肠弯曲杆菌以其诱导人类 DNA 损伤和细胞死亡病理的能力而闻名。这种现象背后的分子机制涉及 Cas9 的核易位，Cas9 是空肠弯曲杆菌 (CjeCas9) 中的一种核酸酶，是 II 型 CRISPR-Cas 系统的分子标记。然而，尚不清楚 CjeCas9 通过哪些细胞途径驱动人类肠上皮细胞进入细胞死亡。在这里，我们展示了空肠弯曲杆菌在感染 Caco-2 人肠上皮细胞期间释放的 CjeCas9 在感染的前四个小时内直接调节 Caco-2 转录组。具体而言，我们的结果显示 CjeCas9 激活 DNA 损伤（p53、ATM（共济失调毛细血管扩张症突变蛋白））、促炎（NF-κB（核因子-κB））信号传导和细胞死亡途径，驱动被野生型空肠弯曲杆菌感染的 Caco-2 细胞，但当被 cas9 缺失突变体感染时，不会导致程序性细胞死亡。这项工作证实了我们之前的发现，即 CjeCas9 具有细胞毒性，并在 RNA 水平上突出了受调节的基础细胞通路。