Research Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
Toxins (Basel). 2024 Sep 19;16(9):407. doi: 10.3390/toxins16090407.
cytolethal distending toxin (CDT) can induce cell cycle arrest and apoptosis. Our laboratory's previous work demonstrated that GTPase 4b (Rab4b) is a key host protein implicated in CDT-induced cytotoxicity. This study investigated the probable involvement of Rab4b in the process. Our study used CRISPR/Cas9 technology to create a Rab4b-knockout cell line. The results showed greater resistance to CDT-induced cell cytotoxicity. In contrast, forced Rab4b overexpression increased CDT-induced cytotoxicity. Further immunoprecipitation study reveals that CDT may bind with Rab4b. In PK-15 cells, CDT is transported to the early endosomes and late endosomes, while after knocking out Rab4b, CDT cannot be transported to the early endosome via vesicles. Rab4b appears essential for CDT-induced cytotoxicity in PK-15 cells.
细胞致死膨胀毒素（CDT）可诱导细胞周期停滞和细胞凋亡。本实验室前期工作表明 G 蛋白 4b（Rab4b）是一种关键的宿主蛋白，参与 CDT 诱导的细胞毒性。本研究探讨了 Rab4b 在这一过程中的可能作用。本研究使用 CRISPR/Cas9 技术构建了 Rab4b 敲除细胞系。结果表明，该细胞系对 CDT 诱导的细胞毒性具有更强的抗性。相反，强制过表达 Rab4b 则增加了 CDT 诱导的细胞毒性。进一步的免疫沉淀研究表明，CDT 可能与 Rab4b 结合。在 PK-15 细胞中，CDT 被转运到早期内体和晚期内体，而敲除 Rab4b 后，CDT 不能通过囊泡转运到早期内体。Rab4b 对于 CDT 诱导的 PK-15 细胞毒性是必需的。
